Abstract: Methods are disclosed for rapid and specific fluorescent staining of biological tissue samples that substantially preserve biological molecules such as mRNA. Methods for microdissecting tissue to obtain pure populations of cells or tissue structures based upon identifying and excising cells or tissue structures that are labeled with fluorescent specific binding agents are also included. A laser capture microdissection apparatus useful for identifying and isolating cells and tissue structures following rapid immunofluorescent staining is also disclosed.

Abstract: A method for producing standard control samples to be used to evaluate disease states or trauma that involve apoptosis or suppression of apoptosis is disclosed. Also disclosed are standard control samples produced by the method. The control samples comprise natural or artificial tissues treated in vitro to display reproducible, predetermined indicators of apoptosis that are equivalent to indicators of apoptotic status of corresponding tissues and organs of a living subject.

Abstract: Prothymosin expression is up-regulated in endometriotic tissue. This invention provides methods of diagnosing endometriosis by detecting up-regulation of a prothymosin gene product, and methods of treating endometriosis by down-regulating expression of prothymosin in ectopic or eutopic endometriotic tissue.

Abstract: Disclosed are methods of identifying psychotropic agents that do not induce motor side effects using differential gene expression. Also disclosed are novel nucleic acid sequences whose expression is differentially regulated by psychotropic agents.

Abstract: A method and apparatus for specimen slide (710) preparation is disclosed. The method and apparatus of the present invention uses slide trays (700) that have receptacles for at least one specimen slide (710) and an associated reagent pack (720). The specimen slide (710) and reagent pack (720) includes respective identifiers (420, 411) that specify a particular slide preparation protocol that should be followed. The method reads the identifiers (420, 411) on the reagent pack and the slide to verify that the correct slide preparation protocol is being used.

Abstract: An apparatus and process for the micro juxtaposition is set forth where a selectively activatable surface is maintained spaced apart from the tissue sample and juxtaposed to the tissue sample by activation. In the typical case, activation occurs by laser radiation with the material of the activatable surface thermally expanding and bringing about the desired micro juxtaposition. The disclosed micro juxtapositioning can cause locally and microscopically pressure on tissue sample, insertion to the tissue sample, or contact of an activated or prepared surface to the tissue sample.

Abstract: An object of the present invention is to provide a method which comprises adhering a biological sample to a film, cutting out a target area of the sample, and then peeling off the film to collect the fragment which was cut out, wherein the operability is good and a cutting sharpness in cutting by a laser beam is good. The present invention provides a method for cutting a biological sample by light irradiation, which comprises locating the biological sample and a colored film having a thickness of 3 to 6 &mgr;m onto one side of a support, and irradiating the biological sample with a light beam, thereby cutting out a target area of the biological sample.

Abstract: The present invention provides the visualization of the fiber architecture in the nervous tissue without staining. The major principle of the method is to make the neural tissue transparent in normal light, and to utilize the ability of neuronal fibers to deflect and deviate light directed from the side to render them visible. The method involves the preparation of thick sections (more than 200 mm) of the nervous tissue, their fixation in paraformaldehyde and dehydration in ethanol. Oil of wintergreen (methyl salicylate) is utilized to make the tissue transparent under normal (bright-field) light. Dark-field illumination is used to create illuminating rays of light arriving at an angle exceeding the collecting angle of the objective lens, thus causing only the axonal pathways to be visible as a bright silver silhouette against a dark background.

Abstract: A histological staining technique that allows quantification of previously unmeasured parameters involved in surgical arteriovenous malformation (AVM) embolization. The invention allows the evaluation of the polymerization characteristics of various ratios of embolization agents, such as Lipiodol/n-butyl 2-cyanoacrylate (NBCA)/glacial acetic acid (GAA) mixtures, by virtue of a new tissue sample preparation protocol and staining technique. To determine the depth of NBCA penetration within the AVM model and to characterize the polymerization patterns of various mixtures within a model vessel, histologic cross-and longitudinal sections were prepared for microscopy using a new staining method including the use of europium aryl-&bgr;-diketone complex and petroleum ether. Paraffin-embedded tissue sections were subjected to the staining protocol to improve differentiation between NBCA and Lipiodol.

Abstract: The present invention relates to prognostic methods which are useful in medicine, particularly cancer chemotherapy. The object of the invention to provide a method for assessing GST-pi expression levels in fixed or fixed and paraffin embedded tissues and prognosticate the probable resistance or sensitivity of a patient's tumor to treatment with platinum-based therapies by examination of the amount of GST-pi mRNA in a patient's tumor cells and comparing it to a predetermined threshold expression level. More specifically, the invention provides to oligonucleotide primer pair GST-pi and methods comprising their use for detecting levels of GST-pi mRNA.

Abstract: The present invention provides reagents for use in an automated environment for cell conditioning of biological samples wherein the cells or tissues are predisposed for access by reagent molecules for histochemical and cytochemical staining procedures. The reagents comprise components optimized to facilitate molecular access to cells and cell constituents within the biological sample. The present invention also provides reagents for use in an automated environment for removing or etching embedding media by exposing a biological sample to be stained in histochemical or cytochemical procedures without the dependence on organic solvents. The reagents comprise components optimized to facilitate removal or etching of the embedding media from the biological sample.

Abstract: A device for the detection of electromagnetic radiation, wherein the device has (i) a photoactive layer of a semiconductor having a band gap of greater than 2.5 eV, (ii) a dye applied to the semiconductor, and (iii) a charge transport layer comprising a hole conductor material, where the hole conductor material is preferably solid and amorphous.

Abstract: The glue-on tissue mount is a device to hold small and/or irregular-shaped biologic tissue so tissue can be manipulated, cut, trimmed, split or divide for transplant and reconstructive surgery. Biologic tissue is quickly mounted onto an acrylic mount with fast-bonding cyanoacrylate glue. On the mount tissue can be held firmly. The tissue can be cut on the mount like on a cutting board or dissected into layers. The mount can rest on flat surfaces and be used with an operating microscope or held between fingers. The glue-on tissue mount has a spherical acrylic mounting surface 12 or flat acrylic mounting surface 14 upon which the tissue is glued. The size and shape of the tissue determines if a spherical or flat mount is used. The mounting surface is attached to a cylindrical base 10 that is easily held. If the glue adhesion breaks the tissue can be anchored with sutures tied to the eyelets in the hexagonal footing of the base 10.

Abstract: Compositions and methods are described for enhancing the immunohistochemical staining of tissue samples. Compositions include a single solution, which includes at least one surfactant, adapted to remove an embedding medium of a tissue sample, rehydrate the tissue sample, and enhance the immunohistochemical staining of the tissue in relation to immunohistochemical staining of unreacted tissue. Methods include heating the tissue sample with the composition to enhance the immunohistochemical staining of the tissue.

Abstract: A monoclonal antibody binding selectively to neurosin obtained from hybridomas, in particular, strain 2B2-6 and strain S2E5 showing stable proliferation ability. These hydridomas are obtained by fusing mouse spleen cells having a high antibody titer against neurosin with mouse-derived myeloma cells, screening fused cells being highly reactive with neurosin, and thus producing an antibody binding specifically to neurosin. By using this antibody, various diseases in which neurosin participates can be diagnosed.

Abstract: This present invention covers novel means, devices and instruments for production of a tissue array block that is further sectioned into duplicates of tissue arrays. An integral microscope is incorporated into the instant instrument for viewing and examining a stained reference slide and selecting donor tissue core region(s) from the reference slide. The said reference slide is held in a reference slide station that is operatively linked and indexed with a station or platform holding a source donor tissue block, which is further indexed and precisely positioned with reference to the donor punch needle for punching the donor tissue core(s). A recipient block indexed to the donor block punch is placed under the donor punch station and donor tissue cores are delivered into pre-existing hole(s) by a stylet to construct the tissue array block. The instant instrument comprises a donor punch station, optionally a second recipient punch station, with each operable independently or removable.

Abstract: Described is an immortalized human skin cell type line (preferably keratinocytes, fibroblasts and/or endothelial cells) stably transfected with a vector, wherein said vector comprises a promoter inducible by environmental hazards and a reporter gene in operative linkage thereto. Preferably, the keratinocyte line is the line HaCaT, the fibroblast line is the line WI-26 and the endothelial cell line is the line LT-2. In addition, a living skin equivalent is described. Furthermore, in vitro toxicity assays making use of these immortalized human skin cell type lines or the skin equivalent are described.

Abstract: Methods are disclosed for rapid, reliable and simple isolation of RNA from formalin-fixed paraffin-embedded tissue samples. RNA purified in this manner can be used for quantitative measurement gene expression levels. The tissue sample can be a tumor or other pathological tissue.

Abstract: Methods are disclosed for rapid, reliable and simple isolation of RNA, DNA and proteins from formalin-fixed paraffin-embedded tissue samples. RNA purified in this manner can be used to monitor gene expression levels. The tissue sample can be a tumor or other pathological tissue.

Abstract: An improved method and apparatus for staining samples of biological material for accurate analysis of the sample. Biological material is applied to a substrate, such as a microscope slide. The biological specimen then is then stained with a selected staining composition, which may be gentian violet for a Gram's Stain analysis. The stained biological material is at least partially decolorized and the level of decolorization is analyzed optically. If necessary, the decolorizing step and the optical analysis steps are repeated until a selected level of decolorization is obtained.

Abstract: Herein is disclosed a method of cross-linking a tissue, comprising treating the tissue under effective cross-linking conditions with a diunsaturated organic compound comprising structure I: wherein R, R′, and R″ are each independently an organic moiety having from 1 to 20 carbon atoms. Also disclosed is a cross-linked biological tissue produced by treating the tissue according to the above method.

Abstract: The present invention relates to toxins that specifically bind to GPI anchored proteins. More specifically, the present invention encompasses the uses of such toxins to detect the presence or absence of GPI anchored proteins. In one embodiment the present invention can be used to detect the presence of paroxysmal nocturnal hemoglobinuria.

Abstract: An improved method of performing immunohistochemical staining on a tissue sample to determine the presence of cytoplasmic tumor marker Metallopanstimulin in cells in the tissue sample. The method consists of generally of collecting the tissue sample; fixing the tissue sample in a manner that preserves the Metallopanstimulin in the cytoplasm of the tissue cells; embedding the sample in paraffin; deparaffinizing the tissue; heating the sample to expose antigenic sites; incubating the slide with a stain blocking agent; incubating the tissue with a primary anti-Peptide A antibody having an affinity for the N-terminal portion of the Metallopanstimulin; incubating the sample with chromogen stain; rinsing the sample; dipping the slide in a counterstain; mounting the slide for reading. Materials for performing the above steps are provided in a convenient, reasonably priced kit.

Abstract: The preparation of phosphorescent metalloporphyrin labelling reagents and their use for preparation of phosphorescent conjugates with biomolecules. The labelling reagents obtainable are water soluble monofunctional derivatives of Pt- and Pd-coproporphyrins, where the term “monofunctional” refers to the number of reactive groups in the porphyrin moiety.

Abstract: A device particularly suited for constructing frozen tissue microarrays. The device comprises: a cooling chamber for receiving at least one frozen material and for maintaining the frozen material in a frozen condition; the cooling chamber moveable in an x and y direction relative to a horizontal surface; at least one coring needle comprising a cutting surface and a lumen for receiving a core of frozen material cut by the cutting surface; and at least one coring needle positioning element, for positioning the at least one coring needle over said frozen material for cutting said frozen material. Frozen tissue microarray blocks and methods of generating these are also provided.

Abstract: The present invention relates to prognostic methods which are useful in medicine, particularly cancer chemotherapy. The object of the invention to provide a method for assessing HER2-neu and/or EGFR expression levels in fixed or fixed and paraffin embedded tissues and prognosticate the probable sensitivity of a patient's tumor to treatment with receptor tyrosine kinase targeted chemotherapy by examination of the amount of HER2-neu and/or EGFR mRNA in a patient's tumor cells and comparing it to a predetermined threshold expression level for those genes. More specifically, the invention provides to oligonucleotide primer pairs EGFR and HER2-neu and methods comprising their use for detecting levels of EGFR and HER2-neu mRNA, respectively.

Abstract: An in vitro model system for viral infection is comprised of a tissue block from adult tonsil or lymph node supported on a matrix which is flexible and porous and wherin the supported tissue block is cultured in a medium whose surface is congruent with the tissue block/matrix interface. The histoculture system can be used to screen for antiviral drugs and to monitor the course of viral diseases.

Abstract: A method is described of viewing a single tissue sample with a light microscope and an electron microscope. A tissue sample is cryopreserved, cryo sectioned and then floated in a cryoprotectant solution on a surface of a glass slide or in a well and is viewed by light microscopy. The step of floating the tissue sample does not destroy said tissue sample for subsequent electron microscope viewing. The same tissue sample is then prepared by conventional ultrastructural processing for viewing by electron microscopy. The data from the light microscope and electron microscope images of selected sites of the same tissue sample can be compared and analyzed.

Abstract: The present invention provides reagents for use in an automated environment for cell conditioning of biological samples wherein the cells or tissues are predisposed for access by reagent molecules for histochemical and cytochemical staining procedures. The reagents include components optimized to facilitate molecular access to cells and cell constituents within the biological sample. The present invention also provides reagents for use in an automated environment for removing or etching embedding media by exposing a biological sample to be stained in histochemical or cytochemical procedures without the dependence on organic solvents. The reagents include components optimized to facilitate removal or etching of the embedding media from the biological sample.

Abstract: Protective solution for use in a fixation method for the paraffin section technique and comprising (i) at least one amino acid and (ii) at least one sugar compound.

Abstract: A highly specific and sensitive technique for exploring cell physiology is disclosed. The method includes laser capture microdissection (LCM) for selecting small clusters of cells of interest from sections of tissue and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) for characterizing simultaneously a broad variety of biological molecules present in the small cluster of cells.

Abstract: The present invention is directed to a fixative composition, the use of the fixative composition in preparing cytological or histological specimens, and a method of preparing particulate matter, such as cytology, hematology and microbiology specimens, for examination by collecting the particulate matter in a uniform layer, preferably a monolayer, and fixing the particles in a composition according to the present invention. The cytological, hematological, microbiological and histological fixative composition of the present invention contains an aldehyde crosslinker, a polyol and a detergent. The method of the present invention for preserving the particulate or histological specimen uses the fixative composition containing an aldehyde crosslinker, a polyol and a detergent.

Abstract: A method of screening a candidate compound for susceptibility to biliary excretion. The method includes the steps of providing a culture of hepatocytes, the culture having at least one bile canaliculus; exposing a candidate compound to the culture; and determining an amount of candidate compound in the at least one bile canaliculus, the amount of candidate compound in the at least one bile canaliculus indicating the susceptibility of the candidate compound to biliary excretion. Optionally, the culture of hepatocytes is a long-term culture in a sandwich configuration. The method is particularly applicable to the screening of multiple candidate compounds in a single effort.

Abstract: The present invention relates to a method of diagnosing, classifying and grading of tumor growths and to determine whether the use of chimeric polioviruses is a proper course for the treatment of the tumors. More particularly, the method is directed to the use of antibodies to a poliovirus receptor (PVR), CD155, to detect the presence of CD155 on tumor cells in various organs, such as: breast, colon, bronchial passage, epithelial lining of the gastrointestinal, upper respiratory and genito-urinary tracts, liver, prostate and the brain.

Abstract: A method for detecting cancer in a patient. The method comprises the steps of introducing labeled antibodies or labeled AFP to a biological sample of the patient so the labeled antibodies or labeled AFP will react with the AFP receptor binding sites in the biological sample. Next there is the step of identifying AFP receptor binding sites in the biological material which are reacted with the labeled antibodies or labeled AFP to determine the presence of cancer.

Abstract: Methods to detect prion or PrP-Sc protein as an indication of transmissible spongiform encephalopathies (TSEs), including preclinical detection of infected live animals, and postmortem detection methods, are described. In one aspect, the invention is directed to a non-invasive diagnostic assay using third eyelid-associated lymphoid tissue. In another aspect, the invention is directed to monoclonal antibodies that specifically bind a conserved epitope of PrP-Sc protein in fixed or frozen treated tissue.

Abstract: An apparatus and method for producing tissue slides is disclosed. The apparatus includes a holding assembly for manipulating the sample block, a blade assembly for preparing a thin section from the sample block, and a transfer roller mechanism for transferring the thin section to a receiving medium. The apparatus further includes a controller that may track the sample block and thin section. The method includes the steps of first locating a sample embedded within a support medium, which may be paraffin or a similar medium. Next, the embedded sample is oriented in such a way that its working surface is presented. This orientation may entail determining the orientation of the embedded sample with respect to the blade that will produce the largest cross-sectional area. Next, a slice of the sample from said embedded sample is removed and subsequently transferred to a suitable receiving medium, which may include a microscope slide.

Abstract: This invention relates to a rapid and efficient method for carrying out enzyme-linked immunosorbent assay for detection of minute quantities of biomolecules such as antigen, antibody etc. This invention particularly relates to microwave mediated immobilization of antigen or antibody on to the activated surface followed by performing subsequent steps of ELISA by controlled microwave irradiation. The invented procedure has dramatically reduced the total time required for ELISA to less than 10 minutes from hours to days. The invented ELISA procedure is rapid, economical, reproducible and simple and can be automated. The invented procedure is useful for carrying out ELISA in clinical diagnostics, molecular biology, agriculture, sericulture, food technology, environmental science, biomedical research and other related fields.

Abstract: A method for screening candidate antimicrobial compounds is described that utilizes a human vaginal xenograft engrafted in a non-human host. The method may be performed by using pathogen inoculated human vaginal xenografts in order to screen a wide range of candidate antimicrobials administered topically or systemically.

Abstract: An aqueous tissue clearing solution for use in making biological tissues transparent is provided. The aqueous tissue clearing solution is selected from the group consisting of dimethyl sulfoxide, diatrizoate acid, ethylenediaminetetraacetic acid, glucamine, &bgr;-nicotinamide adenine dinucleotide phosphate, sodium diatrizoate, and derivatives of polyoxyalkalene. The aqueous tissue clearing solution is used to make tissue transparent for viewing through microscopy.

Abstract: The present invention clarifies and contrasts stain intact biological tissue samples for microscopic analysis. A biological specimen is depigmented and then contacted with a fluorescent dye, wherein the dye stains the specimen nonspecifically. The depigmented, non-specifically stained specimen is equilibrated in a clearing solution, wherein the refractive index of the clearing solution approximates that of the specimen, thereby preparing the biological specimen for visual analysis.

Abstract: A method for preserving cytological specimens for histological or histopathological use. A cytological sample from a biological tissue such as a tumor or a tumor cell line is dehydrated, disbursed and evenly distributed throughout a volume of molten material. The molten material is then drawn into a tubular member such as a pipette and allowed to solidify. Upon solidification, the cylindrical specimen is partially or completely extruded from the tube for further processing, including fixation, dehydration and molten paraffin infiltration and embedding as required for presentation to a sectioning device such as a microtome. Thin circular slices of the cylindrical specimen are removed from the cylindrical specimen and transferred to a slide, fixed and stained as desired. The device and method enable the production of a slide bearing a spatially homogeneous distribution of cytological material for microscopic examination.

Abstract: A method for screening candidate antimicrobial compounds is described that utilizes a human vaginal xenograft engrafted in a non-human host. The method may be performed by using pathogen inoculated human vaginal xenografts in order to screen a wide range of candidate antimicrobials administered topically or systemically.

Abstract: Releasing the embedding medium from an embedded histochemically reactive tissue specimen is provided by contacting the embedded tissue specimen with a releasing composition under conditions sufficient to release a sufficient portion of the embedding medium associated with the histochemically reactive tissue specimen to permit analysis without substantial adverse effect on the histochemical reactivity of the specimen.

Abstract: A method for screening candidate antimicrobial compounds is described that utilizes a human vaginal xenograft engrafted in a non-human host. The method may be performed by using pathogen inoculated human vaginal xenografts in order to screen a wide range of candidate antimicrobials administered topically or systemically.

Abstract: A method of simultaneously staining for various opportunistic pathogens stains tissues sections with Ziehl-Neelsen carbo-fuchsin, Schiff reagent, and a mixture of metanil yellow and stock fast green solution. In particular, the method is useful for staining tissue sections suspected of containing opportunistic pathogens commonly found in AIDS patients, i.e., acid fast mycobacteria, fungus, and Pneumocystis carinii.

Abstract: The genomic structure including the sequence of the intron/exon junctions is disclosed for KVLQT1 and KCNE1 which are genes associated with long QT syndrome. Additional sequence data for the two genes ARE also disclosed. Also disclosed are newly found mutations in KVLQT1 which result in long QT syndrome. The intron/exon junction sequence data allow for the design of primer pairs to amplify and sequence across all of the exons of the two genes. This can be used to screen persons for the presence of mutations which cause long QT syndrome. Assays can be performed to screen persons for the presence of mutations in either the DNA or proteins. The DNA and proteins may also be used in assays to screen for drugs which will be useful in treating or preventing the occurrence of long QT syndrome.

Abstract: Methods are disclosed for rapid, reliable and simple isolation of RNA from formalin-fixed paraffin-embedded tissue samples. RNA purified in this manner can be used to monitor gene expression levels. The tissue sample can be a tumor or other pathological tissue.

Abstract: Laser capture microdissection occurs where the transfer polymer film is placed on a substrate overlying visualized and selected cellular material from a sample for extraction. The transfer polymer film is focally activated (melted) with a pulse brief enough to allow the melted volume to be confined to that polymer directly irradiated. This invention uses brief pulses to reduce the thermal diffusion into surrounding non-irradiated polymer, preventing it from being heated hot enough to melt while providing sufficient heat by direct absorption in the small focal volume directly irradiated by the focused laser beam. This method can be used both in previously disclosed contact LCM, non contact LCM, using either condenser-side (or beam passes through polymer before tissue) or epi-irradiation (or laser passes through tissue before polymer). It can be used in configuration in which laser passes through tissue before polymer with and without an additional rigid substrate.

Abstract: The invention provides methods and materials related to the diagnosis of eosinophil degranulating conditions. Specifically, the invention provides methods and materials that involve visual types of analysis (e.g., microscopic analysis) that are used to determine the presence or absence of a horseshoe-shaped eosinophil granule structure within a mucus sample collected from a mammal. The presence of a horseshoe-shaped eosinophil granule structure within a patient's mucus indicates that the patient has an eosinophil degranulating condition. In addition, the invention provides methods and materials that involve immunological types of analysis (e.g., immunoassays) that are used to determine if a patient's mucus contains a tissue-damaging amount of eosinophil granule content that is outside the eosinophil granule and within the mucus.