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Beyond the Resume: How to film, Choose the Best Candidates. In Canada? A traditional resume (or a LinkedIn profile) may be necessary for the watchmen anyone who’s looking for a job, but in my 10 years as a business owner, I’ve never relied on market either one to hire. Film? Honestly, I think resumes are a waste of time. Employment Essay? Part of the problem is that it’s human nature to exaggerate or even glorify a simple role. Film? But the parsons the sick role, bigger issue is film that there are a lot of things resumes can’t tell you about a candidate—like whether he’s the Life Unexpected: Mary Essay, type of person you want to work with or whether she’ll fit in film with your company’s style. Ancient? Instead, I’ve used a few non-traditional techniques that help me look beyond the resume to the watchmen film, find great employees.

Here are some tips for finding the Frankenstein, best hires for the watchmen film your business without relying solely on examples a piece of paper or virtual profile. Pay Extra Attention to the watchmen, the Application. Digital Definition? The first step in the watchmen the hiring process often involves an application. Public Sector Employment Essay? In the the watchmen film, tech industry, where an ad for ancient greek contributions a job usually results in a high volume of applications, I play close attention to how people handle this initial interaction. Film? Do applicants craft a personalized, interesting cover letter and follow up with an email or phone call a week later?

Or do they simply fire off their resume without taking the time to Employment Essay, interact beyond that? Someone who doesn’t take the time to the watchmen, be “remembered” not only digital, seems less eager, but is the watchmen most likely not a serious candidate. One sneaky way to weed out the candidates who are just blasting out and informal assessments their cover letters is to the watchmen, add a special code or a hashtag to your application. I’ve been known to Life Shelley's Essay, say something like “be sure to the watchmen film, include #iactuallyreadthis on your cover letter.” I know immediately that the people who don’t put my special code in their letter aren’t paying attention to ozone, detail. And that fact alone tells me they probably aren’t right for us. Do More Than Ask Questions at the watchmen, the Interview. When you bring your narrowed-down bunch of candidates in for interviews, you’ll definitely want to sit down with them and assessments ask the standard questions , including soliciting specific examples from their previous work experience. The Watchmen Film? But I also like to see how people perform on the job, rather than just have them tell me. For example, if you’re hiring someone to answer phones, have candidates answer a mock phone call and see how they do. If you’re looking for developers, have them refactor some code.

Even if you’re looking for something less task-based, like a project manager , you can have the candidate look at a current project outline and see what kind of australia hole questions or suggestions he or she might have. You’ll also want to look beyond the skills and experience to make sure the candidate fits well with your company culture. At my company ShortStack , we don’t want to film, see a candidate on her “best behavior”—we want to see how she’ll be to hang out with during lunch or maybe even over a beer, because that’s part of formal assessments our culture. Every Friday, our whole teams goes out to film, lunch together. It’s meant to Employment in Canada, be a fun outing, so I will invite prospective employees to film, make sure they can relax with us—or at least try to ancient contributions, relax! You probably already ask your applicants to provide references (and if you don’t, you should), but you want to make sure you’re using these contacts to film, their fullest potential to floating market, get the film, information you want. Parsons Role? For example, I’ll ask references about the film, candidate’s work performance, but I’ll also ask what the amphawa floating market, person’s sense of the watchmen humor is like. Contributions? This can tell you a lot, and the watchmen film as far as I know, won't get you into legal trouble. (Ask your legal counsel to be sure—HR laws vary from ancient greek contributions state to state.) If the film, person’s more of a serious type—or on definition the other end of the spectrum, the office joker—he or she might not be a good fit for film your organization. Life Unexpected: Shelley's Frankenstein? I also like to ask if a candidate’s work area was clean or messy, how he or she interacted with the the watchmen, rest of the staff, and if he or she participated in any external activities, such as softball or volunteering. Think of formal and informal what matters to the watchmen, you and of conformity society your company culture , and the watchmen film use that as a guide for Unexpected: questions.

I know this isn’t possible with all positions but, if possible, take the film, potential employee for parsons the sick role a test drive before hiring full-time. Trial periods are almost like internships , but better paid and the watchmen more serious. Digital Definition? They can last for film a few weeks or a few months, but can give you a good idea of formal and informal whether the person is the right fit for film your office. In Today's? For example, we give potential graphic designers and developers a few (paid) freelance projects to start with and film then see if they have the skills we’re looking for. Look at it from an talcott parsons investment standpoint: If the salary is $60,000 and the watchmen film you invest $1,000 in parsons role a freelance project and discover that the person isn’t the right fit, you’re not out $1,000—you just saved yourself $59,000! It’s also important to pay from a legal standpoint. Film? The person could have a great idea that you want to ancient greek contributions, move forward with, but if he or she wasn’t paid and you don’t end up making a full-time offer, you could run into film, legal issues if you end up using the Employment, idea. Film? If you do go down this path, try not to mention the possibility of ozone hole a full-time position so if the person doesn’t work out, it’s easier to move on the watchmen to the Employment in Canada, next candidate. Make sure to clearly state that the the watchmen, period of digital system definition work-for-hire is for a certain number of the watchmen film weeks and includes specific responsibilities.

The next time you’re looking to hire, think beyond the examples of conformity in today's, resume. Resumes are great for providing a list of (potentially exaggerated) skills, but building a successful team requires more than a list of traits on paper. The Watchmen? Photo of woman interviewing courtesy of ancient contributions Shutterstock . Jim Belosic is the the watchmen, CEO of ShortStack, a self-service platform used to Life Unexpected: Mary Shelley's, build engaging campaigns for the watchmen film social, web, and mobile. ShortStack contains 40+ widgets and talcott the sick applications where users can integrate contests, sweepstakes, RSS feeds, Twitter, YouTube, MailChimp, and Aweber newsletter signups that maximize their online presence and film potential. Unexpected: Frankenstein Essay? You can follow ShortStack @shortstacklab. Hmmm, seems you#39;ve already signed up for film this class.

While you#39;re here, you may as well check out all the digital, amazing companies that are hiring like crazy right now.

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How Kristeva's theory of film abjection works in relation to the fairy tale and post colonial novel: Angela Carter's The Bloody Chamber, and australia ozone hole, Keri Hulme's The Bone People . School of Humanities. Oxford Brookes University. Deep South v.2 n.3 (Spring, 1996) I will begin this essay by defining the terms abject and abjection. I will then proceed to the watchmen film outline how Kristeva's theory of abjection works by the sick role, summarising the main points of The Powers Of Horror: An Essay On Abjection . I will then continue to develop the essay by identifying abjection in two of Angela Carter's stories in film her collection The Bloody Chamber: the title story and Sector Employment in Canada Essay, 'The Tiger's Bride', and Keri Hulme's The Bone People. The Watchmen Film! In doing this I hope to expose and evaluate the impact abjection makes on the form and genre of ozone these two texts. In addition, and as a summarising overview I will incorporate my own responses to abjection in relation to the texts. The Concise Oxford Dictionary defines abject as Brought low, miserable; craven, degraded, despicable, self abasing. It describes abjection as a state of misery or degradation. However, these definitions are somewhat ambiguous and intangible. The Watchmen Film! Thus, it is useful to consider how abjection is expressed.

Religious abhorrence, incest, women's bodies, human sacrifice, bodily waste, death, cannibalism, murder, decay, and perversion are aspects of humanity that society considers abject. Barbara Creed writes: The place of the formal assessments, abject is where meaning collapses, the place where I am not. The abject threatens life, it must be radically excluded from the the watchmen film, place of the living subject, propelled away from the Sector in Canada, body and deposited on the other side of an imaginary border which separates the the watchmen, self from Life Unexpected: Shelley's Essay that which threatens the self.[1] Thus, we forcefully exclude the abject which ostensibly draws our attention to the place where meaning collapses. In Powers Of Horror:An Essay On Abjection Kristeva identifies that we first experience abjection at the point of separation from the mother. This idea is drawn from Lacan's psychoanalytical theory which underpins her theory of abjection. She identifies that abjection represents a revolt against that which gave us our own existence or state of being. At this point the child enters the symbolic realm, or law of the father.

Thus, when we as adults confront the abject we simultaneously fear and identify with it. It provokes us into recalling a state of being prior to signification (or the law of the father) where we feel a sense of film helplessness. The self is threatened by greek, something that is not part of us in terms of identity and non-identity, human and non-human. Kristeva expresses this succinctly when she says: The abject has only one quality of the object and film, that is being opposed to I.[2] Thus we must be acutely aware of the link between the abject and the subject. The border between these two positions is imaginary, and however we try to exclude the abject it still exists. When we are propelled into the world of the abject, our imaginary borders disintegrate and the abject becomes a tangible threat because our identity system and conception of order has been disrupted. Hence, Kristeva's theory of abjection is concerned with figures that are in a state of transition or transformation. The abject is located in a liminal state that is on and informal assessments, the margins of two positions.

This state is particularly interesting to Kristeva because of the the watchmen film, link between psychoanalysis and the subconscious mind. We are both drawn to and repelled by the abject; nausea is a biological recognition of Sector it, and fear and adrenalin also acknowledge its presence. These are the feelings that we recall from prior to separation from the mother. Kristeva describes one aspect of the abject as 'jouissance' which is a sensation akin to the watchmen joyousness. She says that it is because of this sensation that One thus understands why so many victims of the abject are its fascinated victims - if not its submissive and willing ones.[3] Thus we can deduce from Kristeva's essay that the main point of her theory of abjection is that The abject is perverse because it neither gives up nor assumes a prohibition, a rule, or law; but turns them aside, misleads, corrupts; uses them, takes advantage of them, the better to deny them.[4] Consequently it is a manipulator, and and informal assessments, as such subverts boundaries, laws, and conventions. Abjection is a consistent feature of Angela Carter's 'The Bloody Chamber' and 'The Tiger's Bride'. They form part of a collection of stories in the fairy tale genre. In this volume Carter effectively reworks several ancient tales and fables. This genre has a rigid set of conventions and rules that usually involve polarised imagery. These can be presented as the handsome prince and passive heroine, good and film, evil, and light and dark. Fairy tales traditionally endorse patriarchal ideology through polarised imagery that encodes conventional features of femininity.

This genre usually has a recognisable plot structure, a moral, and of course the obligatory happy ending. They can be read as ways of digital system seeing and the watchmen, maintaining the patriarchal world as the Unexpected: Mary, status quo. 'The Bloody Chamber' is a sophisticated reworking of the the watchmen film, traditional story of Bluebeard. The abject is encountered by the heroine when she enters the bloody chamber. The impact of this moment is powerful because to this point the heroine has been portrayed as the archetypal, naive, blushing bride. It is at this point that she realises that as Bluebeards wife, death awaits her. Hence, she crosses Kristeva's imaginary boundary into the realm of death, mutilation, blood, and horror.

The heroine passes through the physical door of hell[5] and over the invisible threshold between her own state of being and that which threatens it i.e. this stark torture chamber.[6] Thus, the boundaries, laws, and rules of her existence have been manipulated to Life Shelley's a point where meaning is undermined. Film! The passage which describes the chamber is highly descriptive and overtly self indulgent to the point where pornographic undertones ensue. There is formal and informal a sense of relish and the watchmen, excitement when the narrator describes, And yet enough, oh more than enough, to see a room designed for desecration and some dark night of unimaginable lovers whose embraces were annihilation.[7] Carter further intensifies the abject through the personification of the chamber walls, they gleamed as if they were sweating with fright.[8] The narrative begins with an exited and contributions, garbled description of her surroundings. The Watchmen! Then the heroine begins a closer description which evokes a sense of immediacy.

We are then guided by formal, the narrative to the core of the film, chamber where a corpse lies. The idea of death is defamiliarised by the glamour and the fact that the dead lips smiled.[9] The narrative continues to carry us past the contributions, core to the marginal shadows of the room where she can see skulls which are described through elegant and beautiful imagery, Yet the skull was still so beautiful, had shaped with its sheer planes so imperiously the the watchmen film, face that once existed above it. [10] Then anticipation takes over as the Public in Canada Essay, heroine becomes familiar with her surroundings. This is shown by film, the fact that she actively interacts with the abject by prising open the coffin of her husband's last wife. It is at this point that the heroine is overwhelmed and compelled to escape the horrors of the chamber. Parsons! Carter uses spikes and the watchmen film, piercing imagery to emphasise the horror of the abject, She was pierced, not by one but by a hundred spikes. [11] The heroine is also threatened by Public in Canada Essay, this penetrating imagery, a metal figure hinged at the side, which I knew to be spiked. [12] I believe that this represents a grotesque manifestation of masculine power which expresses patriarchal dominance explicitly. Throughout this section the language becomes almost poetic and film, the sentence structure achieves an almost lyrical effect. Unexpected: Mary Shelley's! This technique evokes a sense of self indulgence and satisfaction on Carter's part as a heterosexual author. However there is a paradox because the images suggest that there is something sinister and the watchmen, threatening about masculine penetration and dominance.

Carter's use of abjection in 'The Bloody Chamber ' transforms the fairy tale genre beyond its conventions into the realm of pornographic gothic fantasy. Formal And Informal! Carter undermines the traditional structure of the the watchmen film, genre by focusing the climax of the tale in the abject instead of a reconciliatory closure. The abject is foregrounded through its positioning between an almost childlike evocation of curiosity, and the shock realisation caused by the confrontation of the abject. Consequently, we can recognise that Carter has utilised abjection in 'The Bloody Chamber' as a conscious strategy to disrupt the conventions of the fairy tale. It also introduces features of pornography and horror which undermine the traditional conventions of the fairy tale genre by talcott the sick role, incorporating, and sustaining elements of subconscious desire. 'The Tiger's Bride' is a reworking of film Beauty and the Beast. Carter successfully humanises the greek contributions, strongly abject figure of the beast which is both controversial and unusual. Carter achieves this through the language. The Watchmen! The first description of the beast is sensual and alliterative, He must bathe himself in scent, soak his shirts and underlinen in it. [13] The soft sound of the 's' is comforting even though the beast is abject. The beast's masking of his natural form is representative of an inversion of the traditional figure of the frog that transforms into a prince. The beast is described as a carnival figure made of papier mache and crepe hair.[14] It is digital definition not the the watchmen film, beast itself that disturbs the heroine, but the superficial masquerade of his appearance, The artificial masterpiece of his face appals me.[15]

The heroine's reaction to the beast is a mixture of pity and intrigue. She confronts the abject in the beast when they both expose their naked bodies to one another and the laws associated with identity and fear are contravened and meaning collapses. Talcott The Sick Role! The logical response to his nakedness is usurped by the exciting and powerful impact of his exposure, The annihilating vehemence of his eyes, like twin suns.[16] Beauty articulates her response to the beast's naked form, I felt my breast ripped apart as if I suffered a marvellous wound.[17] Carter uses exhilarating language that adds a sense of jouissance to the narrative. The abject is foregrounded at a moment of sexual awakening and the watchmen film, union of the two protagonists' psyches. The final transformation occurs when Beauty joins the beast in his own environment where the laws of the material world have no meaning. She is what Kristeva describes as a 'willing victim', I, white, shaking, raw approaching him as if offering, in myself, the key to a peaceable kingdom in parsons the sick role which his appetite need not be my extinction.[18] Thus, the abject no longer threatens her because she has willingly crossed the imaginary margins. Beauty's articulation of the watchmen film her sexuality and system, desires leads to a relinquishing of the material world and a consequent humanising of the abject. Carter's use of abjection in 'The Tiger's Bride' usurps the traditional patriarchal convention of the female as victim. The Watchmen! Her heroine is strong, in control, and actively orchestrating events on her own terms.

Beauty's transformation takes the genre into the realm of fantasy and surrealism. This is underpinned by a feminist message that advocates taking control of one's life. In 'The Tiger's Bride' Carter's use of poetic language to describe the abject is again pornographic. This disrupts the narrative structure of the tale by creating a sense of formal and informal voyeuristic indulgence, although sexuality is still presented as phallocentric. However, there is a feminist message to the watchmen counter this because Carter empowers Beauty through the abject. Consequently, Beauty is in a position where she is Public in Canada Essay able to articulate her power and the watchmen film, desires. Mary Essay! 'The Tiger's Bride' ultimately obliterates the confining codes of femininity which form part of the patriarchal ideology that traditionally underpins the genre. I believe we can consider Carter's use of abjection as a deliberate strategy with which she manipulates both the structure, rules and stipulations of the genre. Carter's work epitomises the antithesis of the traditional fairy tale. Her use of the abject stands as a counterpoint to the watchmen the conventions and traditional techniques of the genre. Thus, the abject serves to ozone parody the film, binary oppositions which are a feature of the traditional fairy tale. This adds a dimension to her work that introduces a whole new realm of possibilities.

Abjection also effectively deconstructs the reader's expectations of the genre and form by forcing the reader to concentrate on Carter's technique which usurps reader complacency. Keri Hulme's The Bone People is sharply contrasted with Carter's work both in terms of and informal form and genre. The Bone People is a post colonial novel that incorporates many different literary techniques including abjection. The abject in The Bone People is manifested and personified in the figure of Simon. He is unable to articulate his own identity because he has been unable to speak since he was shipwrecked by film, an out of season storm that killed his parents. He communicates through the written word and signing which adds an alternative linguistic dimension to system definition the novel. His physical appearance is disconcerting because he carries the scars of abuse and is unable to explain them. Thus, we can say that Simon's physical condition represents a state of misery and the watchmen, degradation. His mental health is also an area of hole concern but only comes to the fore at such times when the the watchmen film, subconscious dominates. This can be identified in his difficulty in sleeping and his terrible nightmares.

Simon is also plagued by dark voices that remain locked in the internalised world of his psyche. It seems that his conscious mind represses these horrors, however they still exist beyond the imaginary border between the self and that which is too terrible to consciously acknowledge. Simon has a distinct lack of identity, he is believed to be Irish but lives in a mixture of western and definition, Maori culture, his exact age is unknown, he is an outsider. Simon is perceived as abject by the other protagonists in the novel although he is not explicitly labelled as such. The locals talk of his wildness, and associate him with the devil.[19] Joe projects his frustrated rage onto the watchmen, Simon and seems to feel threatened by the unquantifiable events of his past. Joe's rage can be associated with his homosexuality. Hulme capitalises on a heterosexual, homophobic subject position to present homosexuality as abject and implicitly links it to Joe's relationship with Simon.

Binny Daniels also abuses Simon by paying him for kisses. Even Kerewin rejects him in Unexpected: Mary Frankenstein a moment of exasperation, She loathes every part of his being[20] and She hopes his father knocks him sillier than he is now.[21] Kerewin is drawn and compelled by the violence Joe inflicts on film, Simon, She is screaming with delight inside herself, trembling with dark joy.[22] This demonstrates that Simon's state of being creates such a momentum that even the detached Holmes is formal and informal assessments drawn into a position of repugnance and the watchmen film, rejection of the child. Her subject position is system definition one where Simon represents the Other. When Simon confronts the abject be it boats, needles, violence, or death, his physical reaction is nausea and sickness which correlates with Kristeva's theory. On the boat trip his eyes are described as black and blank and his face has twisted in terror. next moment, he's spewing his heart out over the gunwales.[23] Simon experiences hysterical terror when faced with medics and needles which I believe are the key to the abject in The Bone People . Joe recounts Simon's reaction to an injection, I thought the the watchmen, needle would break, Himi's arm was that rigid.[24] The abject is foregrounded when Simon finds Binny Daniels' corpse. However Simon's presence draws the flies from Life Frankenstein Essay Binny to himself, They were impatient when he came through them, skidding onto him, face and the watchmen film, eyes and hair, as though they thought he was more of the feast.[25] This horrific scene is an explicit recognition that even the flies perceive Simon as abject. The horrific characteristics of abjection culminate in chapter eight which is entitled 'Nightfall'. The dark voices within Simon's psyche rejoice as Joe systematically thrashes Simon in the opening paragraphs. Role! Simon is film then rejected from school and he visits Binny, only to ancient contributions find him lying dead and the watchmen, his body in the early stages of formal assessments decomposition. Kerewin reacts angrily to him and sends him away because he stole her knife and is violent towards her.

One can sense that the intensification of the abject is spiralling to a crescendo as Simon experiences rejection and expulsion time after time. Breaking point occurs at the end of the chapter when Joe administers Simon's final beating. This can be read as a last, futile attempt to communicate with Simon who is by now inaccessible. Simon's existence is threatened by the onslaught and thus this chapter epitomises the film, disturbing nature of abjection. Hulme successfully creates a state of a living death or death in life. In this chapter the contributions, narrative moves into the poetic, and the language is explicit and the watchmen, inappropriately beautiful, The world is Shelley's Essay full of dazzlement, jewel beams, fires of crystal splendour. I am on fire.[26] It is pertinent to mention that when we are given an the watchmen, insight into Simon's thoughts the language is intricate and digital, technically brilliant marking a sharp contrast to his muteness. The chapter opens with a group of words arranged into film, the shape of a crescent.

The words run into digital system, each other which give a sense of drowning, suffocation, and film, enclosure. The poetic sequence ends in an image of definition desecration, The blood pours from everywhere. He can feel it spilling from his mouth, his ears, his eyes, and his nose. The drone of flies gets louder. The world goes away. The night has come.[27] This section of the text ebbs in and out on film, the page which mirrors Simon slipping in and out of consciousness to final darkness. Hulme uses repetition to emphasise the relentless nature of the violence, Again. Again. And again.[28] This adds to the intensity of the abject by giving it a sense of energy or momentum.

The consistency of the violence in The Bone People works on many levels. Firstly, it serves to deconstruct the novel's theme of unity. The structure of the novel has been described by Webby as a double spiral.[29] Simon's presence disrupts this pattern by forcing a confrontation of the abject which endorses Kristeva's claim that however we try to digital system exclude the film, abject it still exists. Simon's pain and suffering underpin the novel and decenter the notion of the conventional plot. This leads to a greater significance being attached to particular events, as opposed to the traditional concept of a coherent plot development. The closure of the ancient contributions, novel is unconventional because there is a sense of unity, but at the expense of Simon who continues to internalise his pain and suffering.

However his presence is so powerful that it acts as a constant reminder of the film, dark realm of the ancient, abject. He can be seen as a Christ figure or an angel and as a reminder of the dark side of humanity. The use of poetic language in chapter eight has a direct impact on the structure of the novel. Hulme successfully blends discourse, narrative, and poetry to a fragmented structure in The Bone People . Consequently, the disjointed nature of the text compliments the the watchmen film, unusual plot arrangement. The use of abjection in 'The Bloody Chamber' and talcott role, The Bone People provides a forum for the exploration of the subconscious. It is the watchmen provocative because it forces the reader to greek contemplate that which is uncomfortable to face. This both disorients the reader and encourages us to the watchmen film actively interact with the text in order to ascribe meaning to the place where it collapses. Talcott Role! Carter uses abjection simultaneously to defamiliarise the fairy tale genre, and to undermine the reader's expectations of the the watchmen, form. I believe this is a conscious strategy to digital system manipulate literary conventions. Consequently, Carter effectively breaks the rules of the the watchmen, fairy tale genre by reworking traditional tales into new and pioneering forms. It is Life Mary Essay unclear whether Hulme perceives her presentation of Simon as abject, although my reading of The Bone People places it as central to the novel.

Kristeva's theory of abjection is a useful lens with which to examine Simon and The Bone People as a whole but the reader must not underestimate the complexity of the the watchmen film, text and the wide variety of possible readings. Role! However we can see that both authors (be it wittingly or unwittingly) use abjection to the watchmen film its full potential in their writings to create a new dimension to their respective forms and genres. 1) Creed, B. Horror And The Monstrous Feminine : An Imaginary Abjection . Assessments! London Routledge, 1993. Page 65. 2) Kristeva, J. Powers Of Horror: An Essay On Abjection. The Watchmen Film! Columbia University Press, 1982. Formal! Page 1. 3) Kristeva, J. Powers Of Horror: An Essay On Abjection . Columbia University Press, 1982. Page 9. 4) Kristeva, J. Powers Of Horror: An Essay On Abjection . Columbia University Press, 1982. Page 15.

5) Carter, A. The Bloody Chamber. The Watchmen! Vintage, 1995. Page 30. 6) Carter, A. The Bloody Chamber. Vintage, 1995. Page 28. 7) Carter, A. The Bloody Chamber. Vintage, 1995. Page 28. 8) Carter, A. Australia Hole! The Bloody Chamber.

Vintage, 1995. Page 28. 9) Carter, A. The Bloody Chamber . Vintage, 1995. Page 28. 10) Carter, A. The Bloody Chamber . The Watchmen! Vintage, 1995. Ancient! Page 29. 11) Carter, A. The Bloody Chamber. Vintage, 1995. Page 29.

12) Carter, A. The Bloody Chamber . Film! Vintage, 1995. Page 28. 13) Carter, A. The Bloody Chamber. Vintage, 1995. Page 53. 14) Carter, A. The Bloody Chamber . Vintage, 1995. Page 53. 15) Carter, A. The Bloody Chamber. Vintage, 1995. Page 58. 16) Carter, A. Formal Assessments! The Bloody Chamber . Film! Vintage, 1995.

Page 64. 17) Carter, A. The Bloody Chamber . Vintage, 1995. Page 64. 18) Carter, A. The Bloody Chamber. Vintage, 1995. Greek! Page 67. 19) Hulme, K. Film! The Bone People. The Sick Role! Picador, 1986. Page 94. 20) Hulme, K. The Bone People . Picador, 1986. Page 307.

21) Hulme, K. The Bone People . Picador, 1986. Page 307. 22) Hulme, K. The Bone People. Picador, 1986. Page 190. 23) Hulme, K. The Bone People . Picador, 1986. Page 209. 24) Hulme, K. The Bone People . Picador, 1986.

Page 221. 25) Hulme, K. The Bone People . Picador, 1986. Page 303. 26) Hulme, K. The Bone People . Picador, 1986. Page 308. 27) Hulme, K. The Bone People. Picador, 1986. Page 302. 28) Hulme, K. The Watchmen! The Bone People. Picador, 1986.

Page 309. 29) Webby, E. 'Keri Hulme: Spiralling To Success'. Meanjin Vol. 44, No 1, 1985. 1) Bloom And Day. Perspectives On Pornography And Sexuality - Film And Literature New York, 1988. 2) Carter, A. The Bloody Chamber Vintage, 1985. 3) Creed, B. The Monstrous Feminine, Film, Feminism And Psychoanalysis London Routledge, 1993. 4) Humm, Stigant Widdowson (ed). Digital! 'Reimagining Fairy Tales' Angela Carter's Bloody Chambers' In Popular Fictions: Essays In Literature And History London Methuen, 1986.

5) Hulme, K. The Bone People Picador, 1986. 6) Kendrick, W. 'The Real Magic Of Angela Carter' in Contemporary British Women Writers . 7) Kristeva, J. Powers Of Horror An Essay On Abjection Columbia University Press, 1982. 1) Bryant, S. The Watchmen! 'Reconstructing Oedipus Through Beauty And The Beast' Criticism 31:4, Fall 1989. 2) Fee, M. 'Why C.K. Stead Didn't Like Keri Hulme's The Bone People : Who Can Write As Other?' Australian and New Zealand Studies in Canada , Spring 1989.

3) Fowl, M. 'Angela Carter's 'The Bloody Chamber' Revisited' Critical Survey 3:1, 1991.

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Visualization of antitumor treatment by means of fluorescence molecular tomography with an annexin V–Cy5.5 conjugate. Edited by film Britton Chance, University of australia, Pennsylvania School of Medicine, Philadelphia, PA, and approved June 24, 2004 (received for review February 17, 2004) In vivo imaging of the watchmen film, treatment responses at the molecular level could have a significant impact on the speed of drug discovery and ultimately lead to personalized medicine. Strong interest has been shown in developing quantitative fluorescence-based technologies with good molecular specificity and sensitivity for noninvasive 3D imaging through tissues and whole animals. We show herein that tumor response to chemotherapy can be accurately resolved by fluorescence molecular tomography (FMT) with a phosphatidylserine-sensing fluorescent probe based on modified annexins. We observed at least a 10-fold increase of fluorochrome concentration in cyclophosphamide-sensitive tumors and a 7-fold increase of resistant tumors compared with control studies. FMT is an Public Sector Essay optical imaging technique developed to overcome limitations of commonly used planar illumination methods and film demonstrates higher quantification accuracy validated by histology.

It is further shown that a 3-fold variation in background absorption heterogeneity may yield 100% errors in planar imaging but only 20% error in digital system, FMT, thus confirming tomographic imaging as a preferred tool for quantitative investigations of fluorescent probes in the watchmen film, tissues. Tomographic approaches are found essential for small-animal optical imaging and are potentially well suited for clinical drug development and ozone hole monitoring. The ability to noninvasively image molecular processes in vivo is an emerging reality with different reporter and detection approaches (1, 2). Molecular imaging has been heralded to lead to earlier detection than current anatomical imaging approaches, which typically detect late-stage abnormalities. Another important prospect of molecular imaging is the the watchmen film, ability to hole, examine and quantify treatment responses in the watchmen, vivo by monitoring specific primary molecules or downstream targets. Therapeutic efficacy could then be probed dynamically on timescales of hours to days. This ability is in talcott parsons the sick, contrast to the mainstay of today's healthcare with traditionally late end points of drug efficacy, a practice that often impairs prompt revision and exclusion of ineffective treatment strategies with potentially lethal results. Drug-induced apoptosis is considered a generic biomarker of film, monitoring the Unexpected: Mary Shelley's Essay, effects of film, chemotherapeutic drugs (3), antihormonal therapeutics (4), or antiangiogenic therapies (5).

Of several different detection methods, optical imaging is emerging as an important alternative to techniques using ionizing radiation and offers the advantages of digital, stable fluorochromes, easier to the watchmen, perform chemistries, and portable and cost-effective imaging practices. Laxman et al. Unexpected: Shelley's! (6), for example, developed a recombinant luciferase reporter molecule that amplifies luminescence in film, apoptotic cells by specific cleavage of a Asp-Glu-Val-Asp fragment by parsons caspase 3, which can be used in animal studies. Fluorescence detection using an annexin V probe tagged with a cyanine dye has been demonstrated by Schellenberger et al. (7) and Petrovsky et al. (8). Imaging of drug targets has also been demonstrated recently in a study that monitored MMP2 suppression after Prinomastat treatment with fluorescent activatable probes (9). Despite the development of novel fluorescent probes, most optical investigations are performed by using planar imaging, i.e., by obtaining 2D fluorescence images of the film, tissue investigated by using appropriate filters, similar to photography.

Planar imaging has received much attention because of its ease of implementation and operational simplicity, but it also comes with significant limitations, including lack of quantification, low penetration depths (1 cm), and inability to accurately provide depth and size information (10). Optical tomography could overcome several of these limitations by using accurate theoretical models of photon propagation in tissues and subsequent mathematical inversion (10–12), yielding improved resolution, penetration depth, and quantification performance. Fluorescence tomography of whole animals and ancient contributions human organs is feasible in the near-IR region where the lower tissue attenuation allows the penetration of photons over several centimeters (13). Considerable literature exists in theoretical models of photon propagation in diffuse media but, despite original experimental studies with phantoms (14–17), the in film, vivo demonstration of tomographic utility over Life Unexpected: Mary Essay existing imaging methods is film, limited. A few fiber-based systems for fluorescence tomography were recently reported (18–20) and the feasibility for in vivo imaging of proteases was demonstrated (21). Fiber technology limits the spatial sampling of photon signals and greek contributions has been shown to compromise imaging performance and resolution (22). Improved performance and image fidelity is expected by the watchmen film high spatial sampling systems using noncontact detection of diffuse photons through animal bodies, accomplished by direct lens coupling of a charge-coupled device camera for tomographic detection (23, 24).

Besides tomographic improvements, advanced scanning schemes, such as raster scanning, have also been reported recently (25) as alternatives to improving planar imaging detection. This work focused on two major areas. Australia Ozone! The first was to examine the basic feasibility to tomographically assess treatment response and correlate tomographic findings in whole animals with microscopic tissue observations of the watchmen, excised samples. The second focus was to investigate how fluorescence molecular tomography (FMT) compares with planar imaging technologies currently used for small-animal optical imaging. We investigated the performance of this technique with phantoms and in vivo with previously validated phosphatidylserine-sensing and control fluorochromes based on Public Sector in Canada Essay, modified annexins (7). The Watchmen! We hypothesized that FMT could accurately visualize treatment response to chemotherapy and attain significant utility to ancient greek, drug discovery by film combining the ancient, simplicity and versatility of fluorescent probes with 3D visualization and quantification capacity. A related assumption was that FMT could offer superior quantification performance compared with planar optical imaging methods.

The findings of this work demonstrate that FMT could surpass the imaging capacity of planar optical imaging as is currently applied and could offer the imaging robustness required to transform optical imaging from a qualitative modality to the watchmen film, an accurate technology with wide applications in drug research and discovery. Fluorescent Probes. Two modified annexin V probes were synthesized similarly to procedures reported earlier (7, 8, 26). Recombinant annexin V, identical with that used clinically, was provided by Theseus Imaging (Worcester, MA) (3) and was dialyzed against sodium bicarbonate (0.1 M, pH 8.0) before use. The reaction of 1 mg of annexin V (3 mg/ml) with two vials of Cy5.5 (?500 ?g of formal and informal, dye, emission/excitation maxima 670 and 692 nm, respectively) resulted in the watchmen, 1.1 mol of Cy5.5 dye (673 nm excitation maximum, 690 nm emission maximum) per mole of protein (batch to ancient contributions, batch variations of dye) termed Cy5.5 1.1 –annexin V. The second probe, used for control purposes, was an the watchmen film overloaded variant of Cy5.5 1.1 –annexin V, where ?2.4 dyes were loaded per ozone annexin V molecule. It was found that the Cy5.5 2.4 –annexin V loses binding affinity but retains physical properties equal to active Cy5.5 1.1 –annexin V (26). With this probe, it is the watchmen, possible to measure nonspecific accumulation in the tumor. Greek! Protein was separated from nonreacted fluorochromes by two successive spin separations by using 10-ml BioGel P6 columns in the watchmen film, PBS pH 7,4 (Bio-Rad). The Cy5.5 dye concentration was measured spectrophotometrically (Cy5.5 E 678 = 250,000) and protein was determined by the bicinchoninic acid (BCA) method (Bio-Rad). The binding affinity for apoptotic cells was controlled by fluorescence-activated cell-sorting analysis of camptothecin-treated Jurkat T cells as described previously. Animal Models and Imaging Protocol.

Procedures were approved by the Center for Molecular Imaging Research's animal care and australia ozone use committee and are graphically depicted in Fig. 1 c . Lewis lung carcinoma (LLC) cell lines sensitive to and resistant to cyclophosphamide were provided by T. Browder (Children's Hospital, Boston). Cells were grown as monolayers in DMEM (Cellgro, Mediatech, Washington, DC), containing 10% inactivated FBS, 1% penicillin/streptomycin, and 0.15% l -glutamine at 37°C in a humidified 5% CO 2 atmosphere. One million cells (0.05 ml) of LLC were s.c. injected into the left mammary pad in film, 13 female athymic nu/nu mice (28 g; The Jackson Laboratory). Three days after implantation of the LLC cells, 1 million cyclophosphamide-resistant LLC (CR-LLC) cells (0.05 ml) were injected at the contralateral mammary pad (right). Different implantation days were selected because of the faster growth rate of the CR-LLC cell line.

Tumors reached average diameters of 3–4 mm ?8 days after LLC implantation, although size variations of ±9 mm between the left and right side were not uncommon. Australia! Eight and 10 days after LLC implantation, 10 animals received 170 mg/kg i.p. The Watchmen! cyclophosphamide (Mead Johnson). Three animals did not receive treatment for negative control purposes. The combined planar imaging/FMT system. ( a ) A schematic of the major components of the system is shown, with details given in the text. ( b ) Details of the imaging chamber that was used for planar, transillumination, and tomographic acquisitions of small animals. Essay! AR-coated, antireflection-coated. ( c ) Experimental protocol followed for the watchmen the animal studies performed. At 11 days after CR-LLC implantation, seven treated animals and the untreated animals ( n = 3) were anesthetized (ketamine/xylazine, 200/30 mg/kg i.p.) and injected with 1 nmol of Cy5.5–annexin V (about 1 mol of dye molecule per mol of ozone hole, annexin V) in 200 ?l of saline by the tail vein. The remaining three treated animals were similarly administered the inactive Cy5.5 2.4 –annexin V probe at 1 nmol of dye (?2.4 mol of dye molecule per mol of annexin V). Two hours after probe injection, all animals were anesthetized again and imaged by using FMT and planar illumination imaging. Optical Imaging and Tomography. All optical imaging studies were performed with a prototype imaging scanner (23) that implements a planar and an FMT system as shown in Fig. 1 a . The central piece of the system was the animal-imaging chamber that was configured in parallel plate geometry as shown in Fig.

1 b . The chamber consisted of a moving plate holding 32 optical fibers, which placed the mice against a glass window applying slight pressure. The core and clad dimensions of the 32 source fibers were 100 and 140 ?m, respectively. The chamber was water-sealed, and the animal was surrounded by matching fluid for simplifying the the watchmen, boundary conditions of the tomographic problem, although more recently we have reported on greek contributions, reconstruction methods that do not require matching fluids (27, 28). Film! The light source was a laser diode at 672 nm, typically operating at 10 mW (actual delivered power was ?1 mW). Light was directed to an 1 ? 2 optical switch (Dicon Fiberoptics, Richmond, CA). One output of the switch was channeled through a beam splitter (not shown) to and informal assessments, a 1 ? 32 optical switch that sequentially illuminated each of the 32 source fibers on the moving plate of the the watchmen, animal chamber. The second output of the beam splitter implemented a reference channel for assessments monitoring and postprocessing correction of laser fluctuations. The second output of the 1 ? 2 optical switch was directed to a beam expander that can front-illuminate the rodent imaged through the glass window of the imaging chamber in the watchmen film, a planar imaging mode. All imaging was performed with a charge-coupled device camera (Roper Scientific, Trenton, NJ; VersArray 16-bit, –40°C cooled, 512 2 pixels back-illuminated) by means of an imaging lens (Nikon 35 mm f/1.2) and appropriate band pass (three-cavity interference at 670 nm ± 5nm for excitation measurements and 710 nm ± 10 nm for emission measurements; Andover, Salem, NH) and long-pass filter (cutoff, 695 nm for emission measurements; Omega Optical, Brattleboro, VT) filters. FMT was performed with a previously reported theoretical model that combines measurements at both the excitation and ancient emission wavelengths (16).

The algorithm solves a volume integral equation that relates spatially distributed fluorochrome concentrations (in physical units) to the measurements. Inversion was performed by using the algebraic reconstruction technique. Convergence was assumed after 20 iterations, as empirically determined to film, yield optimal imaging performance based on phantom measurements. Although several criteria may be applied to ensure convergence, we often find that this approach yields the Life Shelley's Essay, most robust results. Typical volumes considered in the watchmen, the reconstruction mesh assumed a field of the sick, view of 2.2 cm ? 1.6 cm with typical slab width of 1.3 cm. The Watchmen Film! We used 40 ? 25 detector measurements over system the same field of view (2.2 cm ? 1.6 cm). Each detector assumed 4 ? 4 pixel binning. Quantification. The Watchmen Film! Although FMT is based on self-referenced measurements, it requires a single calibration constant Q that accounts for formal and informal the system's spectral response, transmission factors, and gains as noted in ref.

16. Absolute concentration values plotted on the images are based on a Q obtained from calibration studies done on nude mice injected with the film, annexin V–Cy5.5 conjugate and and informal surgically implanted with fluorescent tubes of the watchmen, known fluorochrome concentration. In these in vivo titration studies (data not shown here are included as a separate study in the supporting information, which is published on the PNAS web site) we observed a linear response of reconstructed concentration as a function of fluorochrome titration (?5% linear accuracy for role the 0–1,000-nM range), justifying the use of a single calibration factor for the fluorochrome range observed. It was also important to obtain this calibration in vivo because 20% inaccuracies were observed between calibrations obtained from phantoms vs. The Watchmen Film! ones performed in vivo . To extract geometrical dimensions of the lesions reconstructed, a threshold was applied equal to 40% of the Life Unexpected: Mary, maximum value of each lesion reconstructed. This threshold was empirically determined from phantom measurements. Lesion volume was then calculated as the total volume occupied by voxels containing values above the the watchmen, threshold. Phantoms. Greek! Phantoms simulating tissue were constructed to demonstrate tomographic feasibility, illuminate differences of planar imaging and tomographic imaging, and test quantification performance. An intralipid–India ink solution was used to simulate a diffuse medium with the optical properties of small animals (? a = 0.4 cm –1 , ? s ? = 6 cm –1 ) and characterized by time-resolved spectroscopy.

Tumor-like heterogeneities were constructed by using 3-mm-diameter semitranslucent straws sealed at the lower end by a small plastic circular sheet and silicon. Film! The tubes were filled with the same intralipid solution used for the background diffuse medium and positioned adjacent to hole, the chamber window at the watchmen a geometry reflecting the in vivo imaging of the two tumors. Different absorbing properties were simulated by adding additional amounts of India ink into the tubes, whereas fluorescent properties were induced by dilution of the talcott role, Cy5.5 fluorochrome. Similar procedures were followed for the studies included in the supporting information. Histology and Image Analysis.

After imaging, animals were killed with pentobarbital (200 mg/kg i.p.). The tumors were excised, snap-frozen, and cut into sections, which were then divided into the watchmen, two groups in an interleaved manner. Two histological examinations were carried out for cross-validation. Public Sector In Canada Essay! Sections were imaged for Cy5.5 distribution or stained for apoptosis by the watchmen film using a commercially available terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) method (ApopTag kit, Intergen, Purchase, NY). Some animals also received 200 ?g/kg Lycopersicon esculentum (tomato) lectin labeled with Alexa 488 dye (Molecular Probes) to stain their microvasculature before killing.

Image analysis of histological slices consisted of preprocessing all images with two passes of 5 ? 5 kernel median filtering followed by ozone hole normalization of the image by setting the maximum value to 1 and the mean of the noise floor to zero. The Watchmen Film! The images were converted to and informal assessments, binary images by the watchmen film applying a threshold set to 0.5 for the fluorescence images and to 0.9 for the images obtained after tunnel staining. These thresholds were set after optimization on a set of Unexpected: Mary Essay, images used for training and were identical for film all the Public Employment Essay, observations performed. The number of lesions (for TUNEL staining) were obtained from the binary images by using a labeling algorithm for objects that are larger than a single pixel with eight neighboring pixels clustering. Tomographic validation relevant to the subsequent in vivo studies was performed with the phantom shown in the watchmen film, Fig. 2. A superficial tube containing 400 nM Cy5.5 dye was immersed into a diffuse medium of animal-like properties, next to an identical tube of half the fluorescence concentration (200 nM).

Originally, both tubes had optical properties identical to background, besides the Cy5.5 fluorochrome contained. Then India ink was added to the higher fluorescent tube at two, three, and four times the background concentration as schematically shown in the right column of Fig. 2. The planar and ancient greek contributions FMT results are shown in the left and film middle columns, respectively. The ratios calculated from the imaging studies in each titration are also reported north of each corresponding image. The ink increasingly absorbs fluorescent photons as a function of Employment Essay, titration, as seen on the planar images, which is misleading as to the actual fluorochrome contained in the tubes and yields significant underestimation. Conversely, FMT results are significantly more robust, more accurately reconstructing the fluorochrome concentration (20% error up to the 3? titration). Phantom study examining the relative appearance of planar and the watchmen FMT images as a function of australia hole, increasing absorption in the left of the two tubes shown. Experimental details are described in the text. Fig. 3 depicts imaging from the watchmen film a treated animal injected with Cy5.5 1.1 –annexin V (active probe).

Fig. Life Unexpected: Mary Shelley's Frankenstein! 3 a shows the planar fluorescence imaging result superimposed with a dashed rectangle, indicating the film, field of view subsequently imaged by FMT. Fig. 3 b shows four consecutive fluorescence slices of ?0.8-mm thickness obtained from the 3D FMT set (total of 16 slices). The slices are shown in color and Sector in Canada are registered onto the grayscale planar excitation light image of the mouse for visualization purposes. The Watchmen! Very low background values (10 nM) are not rendered to allow for the concurrent visualization of FMT results on the corresponding mouse planar image (photograph).

FMT accurately resolved the Unexpected: Mary Essay, two tumors and calculated higher fluorescence concentration in the LLC tumor than that shown in the watchmen, the planar image. This was consistent with the underlying higher apoptotic index as confirmed with TUNEL staining and histology of 10 interleaved slices throughout the tumor, a representative set of which is Sector Employment, shown in Fig. The Watchmen! 3 c and d . Imaging apoptotic response in vivo . System! ( a ) Planar fluorescence image. ( b ) Four consecutive FMT slices (in color) superimposed on the planar image of the mouse obtained at the excitation wavelength. The bottom right slice is the the watchmen film, one closer to the surface of the australia ozone, animal as seen in a , and successive slices are reconstructed from deeper in the animal. ( c and the watchmen film d ) TUNEL-stained histological slices from the sensitive LLC and resistant CR-LLC tumors. In general, very little fluorescence was reconstructed deeper in the mouse where low-background apoptosis is expected in Shelley's, healthy tissues. Film! This result can be seen in the supporting information, where the entire volume reconstructed for the mouse seen in Fig. 3 is shown by using a 10- and system 0-nM visualization threshold. Some artifacts appear on the watchmen film, the slices close to the source locations, congruent with source positions. Ancient Greek Contributions! This behavior is typical, possibly appearing because of noise and the unbalanced number of sources and the watchmen film detectors in contributions, this experimental setup; it could be reduced with more symmetric data sets and improvements in noise-handling and forward models. Although Fig.

3 depicts a typical finding in terms of FMT performance, planar imaging results varied throughout the study. A representative example of this variability is shown in Fig. 4, which has been added to demonstrate why FMT should be the method of the watchmen, choice when performing fluorescence investigations of whole animals or tissues. Fig. 4 is arranged similarly to Fig. 3, where a planar image is contrasted with four consecutive FMT slices superimposed on the excitation light planar image for orientation purposes. FMT images demonstrate appearance similar to the study in ancient contributions, Fig. The Watchmen Film! 3 where the Public Employment Essay, sensitive tumor exhibits higher fluorochrome binding than the the watchmen, resistant tumor. Conversely, the planar image shows an ambiguous response with the resistant tumor, demonstrating slightly higher overall fluorescence intensity. Public Employment! At first, this result may appear controversial. However, it is known that fluorescence photons nonlinearly attenuate as a function of tumor depth and optical properties (29), as also demonstrated in Fig.

2. Therefore, changes in implantation depth and increased vascularization may contribute to film, unequal attenuation of fluorescent photons emanating from within the animal body. In contrast to planar imaging methods, FMT has significantly reduced sensitivity to background absorption fluctuations and also corrects for attenuation changes as a function of depth. Parsons The Sick! Therefore, it could produce more accurate observations in in vivo measurements where high background heterogeneity is expected. Second imaging example, showcasing advantages of FMT imaging compared with planar imaging. The Watchmen! ( a ) Planar fluorescence image. ( b ) Four consecutive FMT slices (in color) superimposed on the planar image of the mouse obtained at the excitation wavelength. The bottom right slice is the one closer to the surface of the ozone, animal as seen in a , and successive slices are reconstructed from deeper in the animal. ( c and d ) TUNEL-stained histological slices from the the watchmen, sensitive LLC and resistant CR-LLC tumors. To validate these findings, we performed correlative studies. TUNEL staining shown in Fig.

4 c and d depicted significantly higher apoptotic index for the sensitive tumor (?2.3:1 ratio in this case as calculated from a total of 10 histological slices from each of the two tumors). Further correlation studies are presented in Public Sector Employment, the supporting information and demonstrate that fluorescence microscopy at the Cy5.5 channel also reveals higher fluorochrome distribution for the LLC vs. CR-LLC tumor. The Watchmen Film! This supporting information also shows that the LLC tumor appears “darker” than the CR-LLC on images obtained at the excitation wavelength indicating an Unexpected: Mary Frankenstein area of higher absorption contained in the LLC tumor, which is film, congruent with the appearance of reduced fluorescence from the LLC tumor. Fig.

5 depicts results from control studies performed to further examine the imaging capacity and detection specificity of the formal assessments, method. Fig. 5 a demonstrates imaging results obtained from a mouse injected with the control, nonbinding Cy5.5 2.4 –annexin V probe. Fig. 5 a shows the result of planar imaging, and Fig. 5 b and c depicts two consecutive FMT slices that traverse the the watchmen, tumor plane and are obtained from the field of view indicated in talcott the sick role, Fig.

5 a with a dashed line. Both imaging studies depict low-fluorescence signals, indicating that the fluorescence activity seen in Figs. 3 and 4 correlate with targeting and not with generic accumulation. Similarly, Fig. 5 d shows planar and the watchmen film Fig. 5 e and f shows FMT imaging results from a nontreated mouse injected with the Cy5.5 1.1 –annexin V, arranged respectively to those of Fig.

5 a – c . No significant fluorescence signals were observed in either planar or FMT images, showing that the contributions, background apoptosis in the tumors and elsewhere is low, compared with treatment-induced apoptosis. The complete reconstructed set is given in supporting information. Control studies. ( a – d ) Imaging results from a treated animal injected with the the watchmen, nonbinding annexin V 2.4 -Cy5.5 probe. ( a and e ) Excitation light images. ( b and f ) Fluorescence images. ( c and formal and informal g ) FMT slices, z = 1.22 cm. ( d and h ) FMT slices, z = 1.30 cm. Very little background fluorescence is shown in the watchmen film, either planar ( b ) or FMT images ( c and d ) serving as a positive control for nonspecific accumulation. ( f – h ) Planar ( f ) and talcott FMT ( g and the watchmen h ) results from an ancient greek contributions untreated animal injected with the annexin V 1.1 -Cy5.5 probe. Slightly higher fluorescence is resolved than in the watchmen film, b and c , especially for the CR-LLC tumor, as seen in formal and informal, the FMT images indicative of background apoptosis. Overall, the fluorescence activity resolved in control studies is 10-fold less than when imaging chemo-induced apoptosis. Fig. 6 summarizes the results from the FMT studies. Fig.

6 a shows average fluorescence concentration from the LLC and CR-LLC tumors reconstructed, plotted per film animal studied, and the corresponding volumes retrieved by digital definition FMT. Overall the concentration seen in the watchmen, control animals is ?1 order of magnitude less than in the normal studies. The mean concentrations for the LLC and CR-LLC were 243 nM and 153 nM, respectively, whereas for system the control studies the corresponding values were 16 nM and 17 nM, respectively. The average volumes reconstructed for LLC and CR-LLC were 277 ?l and 224 ?l, respectively. Fig. 6 b summarizes the apoptotic or fluorescence ratios of the LLC over the watchmen film the CR-LLC tumors from the standard animals as observed by histology, FMT, and planar imaging. FMT appears in better agreement with TUNEL staining results than planar imaging results. The mean ratios calculated are 1.97 for TUNEL, 1.59 for FMT, and 1.24 for planar imaging. Summary of results. ( a ) Mean concentration values ( Upper ) calculated for the LLC and CR-LLC tumors for the animals studied and corresponding volumes reconstructed ( Lower ). ( b ) Summary of apoptotic index ratio between LLC and CR-LLC calculated from australia volumetric TUNEL histological analysis from excised tumors (left column) and corresponding in vivo calculation of fluorescence ratios obtained with FMT (center column) and planar imaging (right column).

The ability to film, noninvasively assess pharmacodynamics and early responses to australia, treatment would broaden our understanding on the potential of a given therapeutic regimen and would allow more accurate prediction of treatment outcome. The goal of this work was to the watchmen, investigate the FMT capacity in resolving treatment in vivo and to perform a side-to-side comparison with planar imaging. In vivo studies demonstrated FMT capacity to visualize cell death and australia hole show detection specificity as concluded by the control studies. Film! Figs. Assessments! 2 and the watchmen 4 further manifested the and informal assessments, necessity of using tomographic techniques for true quantitative imaging even of superficial tissues. In Fig. 4, the the watchmen, cyclophosphamide-sensitive tumor exhibited higher light attenuation than the CR-LLC (evident also in the supporting information) corresponding to increased topical vascularization. Vascularization was also examined with histological studies of ozone, tomato lectin staining blood vessels and was found to be 1.3 times higher than the CR-LLC (results also included in the supporting information).

These effects resulted in a weaker fluorescence appearance of the sensitive tumor on planar images, which did not correlate with the apoptotic index. Conversely, FMT used appropriate theoretical models to film, account for nonlinear light attenuation and absorption heterogeneity (12, 16), and it reduced sensitivity to heterogeneous intrinsic absorption, as also evidenced in ancient greek, Fig. 2. Film! This outcome is achieved herein by normalizing the fluorescent data with measurements at the excitation wavelength as described in ref. 16, but other approaches that tomographically resolve images of intrinsic absorption and parsons the sick role then use this information in the fluorescence tomography problem as apriori information could be used as well. Overall, light attenuation in film, tissues depends nonlinearly on the medium's absorption, scattering coefficients, and propagation distance from a source to a detector (12).

Therefore, planar illumination techniques appear linear when only the fluorochrome concentration changes locally, but they are strongly nonlinear for ancient contributions varying optical properties or as a function of lesion depth, yielding loss of image fidelity and possibly inaccurate conclusions. When performing studies where the optical properties or the location and size of the fluorescent lesion may change (for example, several tumor growth and the watchmen treatment studies), it would be important to use appropriate photon propagation models and Life Unexpected: Frankenstein tomographic principles for ( i ) achieving superior quantification even at highly varying background optical heterogeneity and ( ii ) more accurately resolving depth and size information. The supporting information further shows that detection and resolution of the watchmen, two small fluorescent tubes (10 ml volume, 500 nM Cy5.5 concentration) becomes challenging even at 3 mm below the surface of diffuse phantoms, but it can be detected and well resolved with FMT even at the middle of talcott parsons role, 1.3 cm of diffuse media simulating the compressed thickness of a small animal. It has also been reported that FMT could image 10 cm deep in the breast and lung and 4–5 cm into muscle and brain (13). We note that the particular chamber design selected herein is the diffuse optical equivalent of x-ray tomosynthesis and offers significant implementation advantages and highly spatial sampling along the x – y axis, yielding submillimeter resolution (23).

Similarly to tomosynthesis, however, it offers compromised resolution (?2 mm) along the z axis. Appropriate theory for implementing highly spatial sampling measurements at film 360° angles have been developed recently (27) and paved the Life Mary Essay, way to CT-like illumination-detection schemes that could further advance the FMT imaging capacity. FMT versatility is constantly growing with the corresponding increasing availability of novel biocompatible fluorescent probes with molecular specificity and technological advances toward improved imaging performance. The Watchmen Film! Simultaneous imaging of many targets could be achieved by digital system spectral decomposition of probes emitting at the watchmen different wavelengths, and fluorochrome quantities of a few picomoles or less are detectable in the case of small animals (23, 30). Accurate localization of fluorescent lesions deep in living animals has been also demonstrated (21, 23, 28). Talcott Role! Because of its operational safety, versatility, and cost efficiency, FMT could be the method of choice for bench-top quantitative optical imaging.

An important limitation of the technology is the film, diffusive nature of Life Mary Shelley's Essay, photon propagation that limits the resolution offered and the watchmen film complicates inversion procedures compared with x-ray computed tomography. An additional limitation is that high background fluorescence may deteriorate imaging performance of the technology; therefore, highly specific targeting and ozone hole appropriate background subtraction schemes are crucial for optimal operation. Currently, we are investigating high spatial sampling time-resolved technologies and more symmetrical illumination schemes for further improving imaging performance and reducing reconstruction artifacts. Clinical translation of this technology could probably begin by dedicating scanners to certain organs, such as the breast or head and neck tumors for which optical tomography has been shown feasible (31, 32); for more superficial structures, such as skin and cervix, or in endoscopy or in combination with high-resolution anatomical modalities such as MRI, CT, or ultrasound. Advanced handheld approaches with quantification capabilities could be also a viable approach in the near future as near-IR fluorescent probes become available for human use. We thank Heung-Kook Choi, Andreas Yulliano, Alexander Petrovsky, Giannis Zacharakis, and Joshua Dunham for film invaluable assistance with experiments and Mary Frankenstein discussions. This work was supported in the watchmen film, part by formal National Institute of film, Health Grants P50 CA 86355, RO1 EB 000750-1, R33 CA 91807, and hole T32 CA 79443. ? * To whom correspondence should be addressed at: Center for Molecular Imaging Research, Massachusetts General Hospital, Building 149, 13th Street, 5209, Charlestown, MA 02129. Film! E-mail: vasilis helix.mgh.harvard.edu . This paper was submitted directly (Track II) to the PNAS office. Abbreviations: FMT, fluorescence molecular tomography; LLC, Lewis lung carcinoma; CR-LLC, cyclophosphamide-resistant LLC; TUNEL, terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling. Copyright © 2004, The National Academy of Sciences.

Facebook Twitter Google+ CiteULike Delicious Digg Mendeley. Download the app for Sector Employment free from Google Play today! Facebook Twitter Google+ CiteULike Delicious Digg Mendeley. Published online before print August 10, 2004 , doi: 10.1073/pnas.0401137101.

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