Abstract: Cultures of keratinocyte cells are provided which are free from nonautologous fibroblasts and organ extracts, and which have a high speed of cell amplification for a minimum seeding density. Preferably, a culture contains no more than about 10% autologous non-keratinocyte cells such as star-shaped, non-keratinocyte cells and no more than about 1% autologous fibroblasts. The cultures can be cryopreserved in a buffered isotonic medium containing serum and a cryoprotectant. The cultures are produced by a process that does not involve the use of a feeder layer and organ extracts. A culture medium used contains Medium 199, serum, epidermal growth factor, cholera toxin and/or hydrocortisone, and optionally insulin. A substance for wound healing and for cosmetic applications is derived from cultured human keratinocytes. Lyophilized keratinocyte cell cultures or an extract therefrom is used to provide a pharmaceutical composition. Confluent and cohesive keratinocyte sheets are prepared for use in wound healing.

Abstract: Fetal cells may be obtained from amniocentesis, chorionic villus sampling, percutaneous umbilical cord sampling or in vitro fertilization embryos or products of conception, but are preferably from maternal peripheral blood. Fetal cells may be enriched by density gradient centrifugation. Fetal cells may also be enriched by removing maternal cells with an antibody to a cell surface antigen, e.g. anti-CD45, either immobilized or by fluorescence-activated cell sorting. Fetal cells are also distinguishable from maternal cells by staining, e.g. with a labeled antibody to cytokeratin or to fetal hemoglobin, or for fetal hemoglobin by hematoxylin/eosin, or by in situ hybridization to detect one or more fetal mRNAs, e.g., of fetal hemoglobin or fetoprotein. Amplification may be used in conjunction with the in situ hybridization. Fetal cells circulating in maternal blood may be separated by flow cytometry, sorting on their intrinsic light scattering properties.

Abstract: The present inventors have discovered that humans have a gene that encodes a novel protein of the thymosin .beta. family. This novel protein, herein referred to as thymosin .beta.15, has the ability to bind and sequester G-actin, like other members of the thymosin .beta. family, but unlike what is known about other members also directly regulates cell motility in prostatic carcinoma cells. A cDNA of the human thymosin .beta.15 gene (SEQ ID NO: 1) and having the deduced the amino acid sequence (SEQ ID NO: 2) was isolated. The present inventors have shown that enhanced transcripts (mRNA) and expression of the thymosin .beta.15 gene in non-testicular cells has a high correlation to disease state in a number of cancers, such as prostate, lung, melanoma and breast cancer, particularly metastatic cancers. Accordingly, discovering enhanced levels of transcript or gene product in non-testicular tissues can be used in not only a diagnostic manner, but a prognostic manner for particular cancers.

Abstract: Fetal cells may be obtained from amniocentesis, chorionic villus sampling, percutaneous umbilical cord sampling or in vitro fertilization embryos or products of conception, but are preferably from maternal peripheral blood. Fetal cells may be enriched by density gradient centrifugation. Fetal cells may also be enriched by removing maternal cells with an antibody to a cell surface antigen, e.g. anti-CD45, either immobilized or by fluorescence-activated cell sorting. Fetal cells are also distinguishable from maternal cells by staining, e.g. with a labeled antibody to cytokeratin or to fetal hemoglobin, or for fetal hemoglobin by hematoxylin/eosin, or by in situ hybridization to detect one or more fetal mRNAs, e.g., of fetal hemoglobin or fetoprotein. Amplification may be used in conjunction with the in situ hybridization. Fetal cells circulating in maternal blood may be separated by flow cytometry, sorting on their intrinsic light scattering properties.

Abstract: A method and composition for fixing and stabilizing tissues, cells, and cell components such that the antigenic sites and nucleic acids are preserved is provided. The fixative employs a formaldehyde donor that is non-toxic, non-flammable, and that stabilizes the cell with minimal damage to and alteration of the cell morphology. The cell antigenic sites are left intact so that studies with monoclonal antibodies may be conducted. Vaccines and related immunotherapeutic methods utilizing antigens stabilized by the fixative of the present invention are also provided. Also disclosed is a method for developing a positive control for test reagents and for test instrumentation.

Abstract: A method of determining the proliferative status of a carcinoma is disclosed. One obtains a patient sample and then quantitatively analyzes the sample for NGAL gene expression product. The amount of NGAL expression product is compared with a standard curve to determine the S-phase value. The sample can be breast tissue or breast fluid aspirate. Alternatively, blood can by analyzed for this marker to diagnose metastasis.

Abstract: A method of direct extraction of cellular material from a tissue sample which involves: forming an image field of cells of the tissue sample utilizing a microscope, identifying at least one zone of cells of interest from the image field of cells which at least one zone of cells of interest includes different types of cells than adjacent zones of cells, and extracting the at least one zone of cells of interest from the tissue sample. The extracted zone(s) of cells is subjected to analysis. The overall process of identifying, extracting, transporting, and analyzing the extracted zones(s) of cells can be fully automated.

Abstract: Apparatus for processing tissue samples for histological examination comprises a cassette defining a chamber to receive a tissue specimen and having an opening to allow processing fluids to enter and leave the chamber and to allow, eventually, molten wax to enter the chamber for embedding the tissue sample. The apparatus incorporates a temperature-sensitive valve mechanism for closing said opening once the cassette, with the tissue specimen therein, has been immersed in a container of hot molten wax, so that the wax can be retained in the cassette, around the specimen, when the cassette is withdrawn from the molten wax container, until the wax around the specimen has solidified. The temperature sensitive valve mechanism may comprise a shape-memory element.

Abstract: The present invention provides substantially purified cryptdin peptides having a consensus amino acid sequence:X.sub.1 -C-X.sub.2 -C-R-X.sub.3 -C-X.sub.4 -E-X.sub.5 -C-X.sub.6 -C-C-X.sub.7wherein X.sub.1 is 3 to 9 amino acids; X.sub.2 is one amino acid, preferably Y, H or R; X.sub.3 is 2 or 3 amino acids; X.sub.4 is three amino acids; X.sub.5 is five amino acids; X.sub.6 is 6 to 10 amino acids; and X.sub.7 is 0 to 9 amino acids. The invention also provides a substantially purified mouse cryptdin having a consensus amino acid sequence:X.sub.1 -L-X.sub.2 -C-Y-C-R-X.sub.3 -C-K-X.sub.4 -E-X.sub.5 -G-T-C-X.sub.6 -C-C-X.sub.7wherein X.sub.1 is 3 or 4 amino acids, preferably LRD, LSKK (SEQ ID NO: 1) or LRG; X.sub.2 is 1 amino acid, preferably V, L or I; X.sub.3 is 3 amino acids, preferably KGH or *RG, where * is S, T, K, I or A; X.sub.4 is 2 amino acids, preferably GR, RR or RG; X.sub.5 is 3 amino acids, preferably RMN, RVR, RVF HMN or HIN; X.sub.

Abstract: The human MSH2 gene, responsible for hereditary non-polyposis colorectal cancer, was identified by virtue of its homology to the MutS class of genes, which are involved in DNA mismatch repair. The sequence of cDNA clones of the human gene are provided, and the sequence of the gene can be used to demonstrate the existence of germ line mutations in hereditary non-polyposis colorectal cancer (HNPCC) kindreds, as well as in replication error.sup.+ (RER.sup.+) tumor cells.

Abstract: Novel media and methods are disclosed for the in vitro formation of a histologically complete human epithelium. The media are serum-free, companion cell or feeder layer free and organotypic, matrix free solutions for the isolation and cultivation of clonally competent basal epithelial cells. The media and methods of the invention are useful in the production of epithelial tissues such as epidermis, cornea, gingiva and ureter.

Abstract: The invention relates to methods for determining metastatic potential of tumors and to methods and compositions for inhibiting or preventing metastasis of cancers. In one aspect, the invention provides a method to determine metastatic potential of tumors, particularly prostatic tumors. In this regard the invention relates to determining protein or mRNA of effectors of arachadonic acid release to gauge metastatic potential, particularly uteroglobin protein or mRNA to determine metastatic potential of prostatic tumors.The invention also relates to methods and compositions that prevent or inhibit metastasis of cancers. In this regard, the invention particularly relates to methods and compositions that inhibit arachidonic acid, those that inhibit phospholipase A.sub.2. More particularly in this regard, the invention relates to uteroglobin or muteins, peptide analogs or mimetics of uteroglobin and lipocortins or muteins, peptide analogs, or mimetics of lipocortins that inhibit metastasis.

Abstract: A composition which comprises human mesenchymal stem cells which have the potential to differentiate into cells of more than one connective tissue type and a composition which induces cells from the mesenchymal stem cell population to differentiate into the adipogenic lineage, and a process for inducing such differentiation. The composition for inducing such differentiation comprises a glucocorticoid and a compound which stimulates cAMP production or inhibits cAMP degradation (such as a phosphodiesterase inhibitor). The process can further include isolating the adipocytes from remaining hMSCs.

Abstract: A composition and treatment method for fixation cells and for preparing them for examination. In particular, the invention contemplates compositions effective in inhibiting crystal formation in the cell sample and effective in enhancing cell adherence to examination slides.

Abstract: A method of staining cellular material where a sample containing the cellular material is stained with a stain solution containing a cyanine dye excitable by infrared rays to contrast its nuclear material from its cytoplasmic material.

Abstract: A method and an allele-specific in situ reverse transcriptase polymerase chain reaction (RT-PCR) amplification assay for detection and differentiation between expression of unmutated wild-type DNA sequences and between endogenous mutated DNA sequences in vitro or in vivo. The method is useful for verification, diagnostic assessment and monitoring of therapeutic small fragment homologous replacement gene therapy, for diagnostic assessment and monitoring of cDNA-based gene therapies, for analysis of gene expression of specific alleles during fetal development and for diagnostic assessment of the expression of alleles involved in cancer mutations.

Abstract: The present invention is a method for rendering cotton fiber cells that are post-anthesis and pre-harvest available for analysis of their physical properties. The method includes the steps of hydrolyzing cotton fiber cells and separating cotton fiber cells from cotton ovules thereby rendering the cells available for analysis. The analysis of the fiber cells is through any suitable means, e.g., visual inspection. Visual inspection of the cells can be accomplished by placing the cells under an instrument for detection, such as microscope or other means.

Abstract: Anti-p185.sup.HER-2/neu antibodies which are useful in the detection of HER-2/neu oncogene overexpression in biological samples are described. The antibodies are accurate and reliable in immunocytochemical or immunohistochemical assays of cell and tissue samples. Also described are methods for detecting HER-2/neu oncogene expression in a biological sample using the antibodies of the invention and a diagnostic kit comprising the antibodies. The reagents provide an accurate means of identifying certain cancer patients who have the greatest probability of relapse and/or the least likelihood of survival.

Abstract: Disclosed is a synthetic medium with PDGF, vitronectin, IL-1.beta. and BSA added to Eagle's minimum essential medium or medium with transferrin, insulin, progesterone and putrescine further added thereto. When cultivating the postnatal central neurons using the inventive medium, there are effects such that good attachment to substrate, extension of neuritic processes and maintenance of survival are achieved, that more stable sure cultivation becomes possible as well over the astrocyte-conditioned medium used hitherto, and the like.

Abstract: An assay for determining thrombin receptor activation in a cell by measuring the internalization of a labeled antibody-thrombin receptor complex following exposure to an agonist peptide is provided. This method is useful in identifying thrombin receptor antagonists.

Abstract: The present invention relates to a growth factor preparation derived from a mixed lymphocyte culture, and a process for its production. The invention also relates to a cell culture medium containing said growth factor preparation. The invention further relates to a process for culturing plasma cells and producing antibodies by using said cell culture medium.

Abstract: The invention features a method for the selection and expansion of bone marrow stromal cells. The method includes the steps of obtaining bone marrow stromal cells; introducing the stromal cells into a vessel pre-coated on an inner surface with a gelatin, and containing a culture medium including an acidic fibroblast growth factor ("aFGF") polypeptide; and expanding the stromal cells in the culture medium under conditions and for a time sufficient to obtain an increased number of bone marrow stromal cells. The culture medium additionally can include heparin, and the vessel additionally can be precoated with fetal bovine serum.

Abstract: Fetal cells may be obtained from amniocentesis, chorionic villus sampling, percutaneous umbilical cord sampling or in vitro fertilization embryos or products of conception, but are preferably from maternal peripheral blood. Fetal cells may be enriched by density gradient centrifugation. Fetal cells may also be enriched by removing maternal cells with an antibody to a cell surface antigen, e.g. anti-CD45, either immobilized or by fluorescence-activated cell sorting. Fetal cells are also distinguishable from maternal cells by staining, e.g. with a labeled antibody to cytokeratin or to fetal hemoglobin, or for fetal hemoglobin by hematoxylin/eosin, or by in situ hybridization to detect one or more fetal mRNAs, e.g., of fetal hemoglobin or fetoprotein. Amplification may be used in conjunction with the in situ hybridization. Fetal cells circulating in maternal blood may be separated by flow cytometry, sorting on their intrinsic light scattering properties.

Abstract: A method of assaying leukocyte binding to a tissue sample, which comprises contacting a arterial tissue sample at a temperature of at least 10.degree. C. with a suspension of monocytes or a cell line having adhesion to arterial tissue similar to monocytes, and quantitating the number of bound cells over a defined area of tissue sample section. The assay allows agents which inhibit binding of monocytes to human vascular tissue to be identified, and the use of such agents is described in the therapy of atherosclerosis.

Abstract: A serum-free medium which supports the growth and proliferation of bone marrow cells is described. Recipes for two formulations are given, one of which provides a medium suitable for growth of bone marrow cells for use in human therapeutic protocols.

Abstract: Oligonucleotides that are capable of passive diffusion across cell membranes are disclosed. These oligonucleotides contain at least two nucleotide residues and show a log distribution coefficient in octanol:water of about 0.0-2.5 and a solubility in water of at least 0.001 .mu.g/mL. In preferred embodiments, either at least 80% of the internucleotide linkages are non-ionic, or at least 80% of the bases contain lipophilic hydrocarbyl substitutions, or a combination of these sums to 80%. These oligonucleotides may be conjugated to label and used to visualize cells.

Abstract: A fluorescence in situ hybridization (FISH) procedure and solutions is provided. The entire FISH procedure is fast (15 minutes or less), with the hybridization step occurring in 5 minutes or less. The entire FISH procedure with digoxygenin- or biotin-labeled probes takes approximately 30 minutes. A formamide-free solution (dextran sulfate and glycerol) is provided. An additional solution (10% dextran sulfate and 20% formamide) is also provided. Additional solutions are provided for mRNA in situ hybridization whereby the process takes less than 24 hours.

Abstract: A method for producing cellular fibronectin, said method comprising incubating fibronectin-producing cells in a fibronectin production medium comprising: (a) 1.0-3.5 g/L bicarbonate salt; (b) 1.0-5.0 g/L glucose; (c) 10-30 .mu.g/L dexamethasone; (d) 1-10 g/L hydrolyzed protein; and (e) 5-15 .mu.g/L insulin.

Abstract: The use of individual or combinations of cytokines, particularly IL-3, GM-CSF, and c-kit ligand are employed for long-term hematopoiesis in serum free culture in the absence of stromal cells. The cultures can be used for evaluating compounds and their effect on hematopoiesis, particularly as to lifetime and nature of differentiation. In addition, the expanded cells may be used for engraftment in a mammalian host or enhancement of particular cell lineages in a mammalian host. The subject systems may be used with any mammalian hemopoietic cells, but finds particular application with primates, more particularly humans.

Abstract: Provided is a polymer film useful in the delivery of biological agents, as well as a method for making such a polymer film. The polymer film comprises a transparent, gelled water-soluble polymer supported by a mesh. The polymer film also contains a biological agent. The polymer film can be made by preparing an aqueous solution of a gellable water-soluble polymer and the biological agent, dipping the meshed support into the aqueous solution to create a liquid film over the support, and then gelling the polymer.

Abstract: A method of treating cancer by the use of a multidrug chemotherapeutic regimen determined by in vitro pharmacosensitivity tests. A cell suspension is prepared from a tumor specimen obtained from the patient. The viable tumor cell count within the cell suspension is calculated. The volume of the cell suspension is then adjusted to obtain a base cell concentration by diluting the cell suspension with patient medium in proportion with the viable tumor cell count. A sample of the cell suspension is retained as a negative control sample. Drug samples are then prepared, each drug sample containing a mixture of cell suspension, patient medium, and a drug selected from several drugs, wherein each drug sample contains a different drug which is added to the drug sample in an aliquot amount proportional to the base cell concentration. The drug samples and negative control sample are then incubated. After incubation, the drug samples and negative control sample are stained with a DNA intercalating dye.

Abstract: A novel bronchial or bronchiolar epithelial cell from normal neonatal mammalian lung has been isolated, established and maintained for multiple passages in the absence of serum, without undergoing crisis or senescence. By careful manipulation of the nutritional/hormonal microenvironment we have been able to select, from a heterogeneous population, a single epithelial cell type which can maintain highly differentiated features in vitro. This cell type has characteristics of bronchiolar epithelial cells. A clonal line, RL-65, has been selected and observed for more than 3 years in continuous culture. It has been characterized by ultrastructural, morphological and biochemical criteria.

Abstract: A machine-based method for collecting and interpreting quantitative data on cells and tissues so that a diagnosis will obtain as to the existence or non-existence of disease in an human. Vibrational spectroscopy is used and the spectra generated by such spectroscopy are compared with stored spectra to provide whether cells or tissues are diseased, and if diseased to what degree. It, therefore, is possible to provide a basis for immediate diagnostic decisions for patients and physicians, leading in turn to immediate implementation of next-step procedures and treatment all in one visit to the doctor's office. This means that patients and the examining clinician can know almost instantly whether or not the cells or tissue examined are normal or diseased, and the level of disease present if found.

Abstract: Non-carboxylated sulfated polyanions have been successfully used to rapidly obtain and maintain stable single-cell suspension of BTI-TN5B1-4 cells, a cell line which has a high intrinsic capacity for the expression of recombinant protein, but which clumps severely in suspension reducing its effectiveness as a host for foreign protein production with the baculovirus expression vector system. The three most effective polyanions for inducing a single-cell suspension were dextran sulfate, polyvinyl sulfate, and pentosan sulfate. The cost of dextran sulfate treatment is low compared to heparin treatment, which required a 20-fold higher lever to induce single-cell suspension. More importantly, dextran sulfate does not block vital infection at MOI.gtoreq.1 whereas heparin is known to seriously inhibit infection.

Abstract: The invention relates to a method of detection, a sensor and a test-kit which find application in immunological detection (e.g., immunoassay). The invention provides, inter alia, a method of detection, suitable for use in immunological detection of an entity, which method includes the use of a secondary species (as defined in the specification), the use of a first detectable species, and the use of a second detectable species. The method may include, for example, the use of a primary species, a secondary species, a first detectable species and a second detectable species. The primary species may be, for example, an antibody or a ligand. The secondary species may be, for example, an auxiliary species such as an auxiliary binder or an auxiliary ligand, or a species which has a part which is an auxiliary function. The entity to be detected may be an analyte species as such or may be an entity which carries or includes analytes species.

Abstract: A method for detecting presence or absence of a nucleic acid of interest in fetal nucleic acid derived from a sample of peripheral blood obtained from a pregnant woman is described. The method involves obtaining a sample of peripheral blood from a pregnant woman, treating the sample of peripheral blood such that fetal nucleic acid present in fetal granulocytes is made available for detection and detecting presence or absence of a nucleic acid of interest in the available fetal nucleic acid. The proportion of fetal granulocytes present in the sample of peripheral blood can be increased relative to the sample of peripheral blood forming a sample enriched in fetal granulocytes prior to the detection step.

Abstract: A method for enhancing the survival and/or proliferation of Schwann cells (especially human Schwann cells) in cell culture is disclosed which involves culturing the cells in serum free culture medium comprising gas6 and other mitogenic agents, such as heregulin and forskolin. The culturing step is generally preceded by a pre-incubation period wherein nerve tissue comprising the Schwann cells is cultured under appropriate conditions and for a period of time such that demyelination occurs. The isolated Schwann cells can be used as cellular prostheses to treat patients with nervous system injuries. The invention also provides a cell culture medium for culturing Schwann cells.

Abstract: The present invention provides systems, methods and chemically defined media for the cultivation of cells, particularly epithelial cells. Cells may be cultured with a varied calcium concentration. Furthermore, a calcium concentation in excess of 1.00 mM may be used in the practice of the present invention without loss of a proliferative cell population and maintianing high colony forming efficiencies. Population doubling times range from about 16 to about 33 hours. Cells may serially cultivated to achieve from about 20 to about 50 population doublings.

Abstract: Methods for obtaining cells that produce a ligand for an orphan receptor and methods for preparing polynucleotide molecules that encode ligands for orphan receptors are disclosed. The methods utilize growth factor-dependent parent cells that are transfected with a DNA construct encoding an orphan receptor. The transfected cells are exposed to mutagenizing conditions, and the mutagenized cells are cultured under conditions in which cell survival is dependent upon autocrine growth factor production. Progeny cells are recovered and screened to identify those that produce a ligand for the orphan receptor. Polynucleotide molecules encoding the ligand can be prepared from the identified cells.

Abstract: The present invention is directed to a new preservation solution useful for the initial flushing and for the storage of organs intended for transplantation using a warm preservation technology, between 18.degree. C. and 37.degree. C. Among the components of the preservation solution are a basal mammalian cell culture medium comprising one or more serum proteins, growth factors, particularly retinal-derived growth factor mucopolysaccharides, and emulsified liquid fluorocarbons, and cyclodextrin.

Abstract: A tissue fixative comprising a solvent including one or more alkanols, one or more diols and/or triols and a catalyst, and a solute including an osmotically active substance and a mordant. A preferred composition comprises 14 ml ethanol, 14 ml ethandiol, 2.43 ml of a 37% solution of methanal, 0.375 g NaCl, 0.0375 g ZaCl.sub.2 and 69.57 mls water.

Abstract: A method of determining whether a test cell from a given human tissue type is (a) normal, or (b) cancerous or precancerous, by contacting the mRNA of the test cell with a nucleic acid probe which contains a nucleotide sequence at least 15 nucleotides in length which is complementary to a portion of the coding sequence of a candidate tumor suppressor gene, which gene is one that is expressed at a given control level in normal cells of that tissue type; and determining the approximate amount of hybridization of the probe to the mRNA of the test cell, an amount of hybridization one-third or less that seen with the mRNA of a normal cell of that tissue type being an indication that the test cell is cancerous or precancerous. Alternatively, an antibody specific for the candidate tumor suppressor gene product can be substituted for the nucleic acid probe as a means for determining the level of expression of the gene in the test cell.

Abstract: The present invention describes a method of staining mitochondria, and analyzing mitochondrial function, using a class of fluorescent substituted 3',6'-diaminoxanthenes and their reduced analogs 3',6'-diaminodihydroxanthenes, which are oxidized to the fluorescent form of the dye in situ. In their oxidized form, the dyes selectively localize within mitochondria. The dyes of the invention are substituted by an alkylating group that allows their retention in mitochondria even after cell death, fixation, and permeabilization.

Abstract: Methods and formulations are disclosed for the in vitro formation of a histologically complete human epidermis in a serum-free, companion cell or cell feeder layer-free, and organotypic matrix-free culture system commencing with the isolation and cultivation of a unique population of clonally-competent basal epidermal cells and ending with the formation of a functional, histologically complete, human squamous epithelium. The formation of a histologically complete human epidermis is accomplished in a serum-free medium, without companion-cells or feeder layer cells or any organotypic support using a multi-step process that is controlled by manipulating the growth and differentiation factors requisite to the sequential development of a usable, functional, and completely differentiated epidermis.

Abstract: The instant invention provides for monoclomal antibody 369.2B which is specific for the .beta.A4 peptide, and in particular the free C-terminus of .beta.A4 "1-42" but not "1-43", and stains diffuse and fibrillar amyloid, vascular amyloid, and neurofibrillary tangles. The instant invention further provides for antibody fragments and constructs thereof which have the same binding specificity. The instant invention also provides for methods of diagnosis, screening and therapeutics for treating unique forms of .beta.A4 peptide, using the antibodies of the instant invention.

Abstract: Disclosed is a method of fluorescent detection of a nucleic acid. The method comprises contacting to the nucleic acid a bis-dicationic aryl furan and exposing the nucleic acid to light at a frequency to induce fluorescence of the compound. A method for fluorescent detection of cytoskeleton elements, and novel bis-dicationic aryl furan compounds are also disclosed.

Abstract: Process for rapid, ultrasensitive and automatic counting of fluorescent biological cells such as microorganims, carried by a solid support such as a filter.The process includes: scanning a solid support on which a specimen potentially containing microorganisms has been deposited, with an incident beam from a laser, forming a laser spot on the solid support, the laser spot being substantially greater than the microorganisms to be detected, the laser spot size being between 4 and 14 .mu.m and simultaneously: detecting the resultant fluorescent light at least at one wavelength; establishing a set of correlated-features by a line-to-line correlation of individual features; comparing said correlated-features on each pair of adjacent lines in time synchrony, at least at two different wavelengths .lambda..sub.1 and .lambda..sub.

Abstract: Assays for target molecules in and from cells and viruses, e.g. nucleic acids, wherein non-specific background is decreased by including an analogue of the reporter group, e.g. a non-fluorescent analogue such as fuchsin, of a fluorescent group such as fluorescein, to decrease non-specific background.

Abstract: The invention features a method of labeling a cell containing an intracytoplasmic target molecule involving (1) permeabilizing the plasma membrane of the cell so that (a) a reagent capable of detectably labeling the intracytoplasmic target molecule can traverse the plasma membrane into the cytoplasm of the cell; and (b) substantially all of the intracytoplasmic target molecule and the DNA of the cell remain in the cell; and (2) contacting the cell with the reagent to label the intracytoplasmic target molecule. The method may further involve detecting the label in the cell, and isolating the cell on the basis of detecting the label in the cell. The invention also includes cells permeabilized using the method of the invention.

Abstract: The invention concerns diazapentalene derivatives of formula I ##STR1## in which X and Y represent leaving groups which are less nucleophilic than thiol compounds, as thiol-specific fluorochromes for the detection of compounds containing thiol groups, in particular in cells.