Abstract: Provided are a fluorescent microsphere comprised of a plurality of fluorescent nanocrystals embedded in a polymeric microsphere; a kit prepared from the fluorescent microspheres; and a method of producing the fluorescent microspheres comprising swelling the polymeric microsphere so that into its pores can enter fluorescent nanocrystals, and then unswelling the polymeric micropsheres so that the fluorescent nanocrystals become physically entrapped in the pores of the unswelled polymeric microsphere. Also provided is a method of using the fluorescent microspheres comprising affinity ligand for determining the presence or absence of a predetermined number of analytes in a sample by contacting the sample with the fluorescent microspheres, and detecting the fluorescence signal pattern of excited fluorescent microspheres bound to one or more analytes of the predetermined number of analytes, if present in the sample.

Abstract: An automated cell management system which can be programmed to perform and control various operations of the essential phases of cell culturing, of cell culture manipulation, and of cell culture evaluation. The automated cell management system comprises a housing, a storage array for accommodating a plurality of cell culture devices, a loading station, a means for harvesting one or more components from a cell culture device, and one or more processing stations. The automated cell management system may further comprise mechanism for tracking each cell culture device, a plurality of reservoirs, a centrifuge, a microprocessor, one or more evaluation stations, one or more means for sterilization, and a combination thereof.

Abstract: Provided is a cell culture apparatus for culturing cells, and optionally, for performing magnetic separation of cells desired to be cultured. The cell culture apparatus preferably comprises a frame; at least one gas permeable membrane securedly sealed in a leak-proof sealing to a frame and an opposing surface comprising a rigid surface in forming a culture chamber therebetween; and at least one resealable aperture to allow substances to be introduced into, or withdrawn from, the culture chamber.

Abstract: Provided is a device comprising a plurality of microchambers having a closed vented environment, wherein each microchamber is in operative communication with a filling port and a vent aperture. The device further comprises a base which is sandwiched between two liquid-impermeable membranes, with at least one of the membranes being gas permeable. Also provided is a method for introducing a fluid into a plurality of microchambers of the device, wherein each filling port is aligned with a pipette tip, and the fluid is introduced into and through the filling port. The fluid then flows along a fluid flow groove providing fluid flow communication between the filling port and the microchamber, and into the microchamber.

Abstract: An apparatus for use in a swinging bucket centrifuge rotor, the apparatus comprising a pair of sidewalls wherein each sidewall comprises a plurality of longitudinally spaced and laterally disposed partitions; wherein each pair of adjacent partitions defines a slot which is dimensioned to receive a cell culture device. The apparatus comprises a centrifuge bucket or an adaptor for use with a centrifuge bucket. Also provided is a method for harvesting one or more components of a cell culture contained within a cell culture device by centrifuging the cell culture device as supported by the apparatus during centrifugation, and then removing separately the one or more components of the cell culture from the centrifuged cell culture device.

Abstract: Provided is a cell culture apparatus for culturing cells, and optionally, for performing magnetic separation of cells desired to be cultured. The cell culture apparatus preferably comprises a frame; two membranes, preferably each being gas permeable, which are securedly sealed in a leak-proof selaing to a frame in forming a culture chamber there-between; and at least one resealable aperture to allow substances to be introduced into, or withdrawn from, the culture chamber.

Abstract: Provided is a method for loading dendritic cells ex vivo which utilizes immune complex-mediated uptake of a soluble antigen by dendritic cells. The method involves coculturing target cells in the presence of, but not in physical contact with, expanded dendritic cells. Added to the coculture is human antibody against the soluble antigen(s), resulting in formation of immune complexes. The immune complexes bind to, and are uptaken by, the expanded dendritic cells; and with subsequent processing of the antigen into peptides, the peptide is presented as part of a peptide-MHC complex on the dendritic cell surface. The method of the present invention allows for dendritic cell presentation by MHCI and/or MHCII complexes, depending on the nature of the one or more antigens.

Abstract: Provided are compositions and assay kits comprising functionalized nanocrystals having extending therefrom a plurality of polynucleotide strands of known sequence; wherein primary dots are used to operably link to a molecular probe, and secondary dots comprise a plurality of polynucleotide strands which are complemetary to the plurality of polynucleotide strands of the primary dots. Also provided is a method for detecting the presence or absence of target molecule in a sample comprising operably linking primary dots to molecular probe, contacting the complex formed with the sample, contacting the sample with successive additions of secondary dots and primary dots. If target molecule is present in the sample, the primary dots and secondary dots will form dendrimers that can be detected by fluorescence emission.

Abstract: Provided is a cell culture apparatus for culturing cells, and optionally, for performing magnetic separation of cells desired to be cultured. The cell culture apparatus preferably comprises a frame; two membranes, preferably each being gas permeable, which are securedly sealed in a leak-proof selaing to a frame in forming a culture chamber therebetween; and at least one resealable aperture to allow substances to be introduced into, or withdrawn from, the culture chamber.

Abstract: An in vitro system for perfusion of cultured cells comprising one or more cell culture devices, one or more reservoirs, and a perfusion mechanism for providing a fluid flow between the one or more cell culture devices and the one or more reservoirs. Also provided is a method of using the in vitro system to contact cultured cells, contained in the one or more cell culture devices, with a biological substance. The biological substance may be mixed with medium, and then the medium may be circulated through the one or more cell culture devices so that the biological substance contacts cultured cells contained in the one or more cell culture devices.

Abstract: A method is provided for immune corrective surgical removal of lymphoid tissues containing shed tumor antigen in an individual. A detector molecule, with binding specificity for shed tumor antigen, is administered to an individual, and detected in the individual is the presence of lymphoid tissue which contains shed tumor antigen, as detected by the signal emitted by the detector molecule bound to shed tumor antigen. The lymphoid tissue, detected as containing shed tumor antigen, may then be surgically removed, thereby removing B cells, shed tumor antigen, and follicular dendritic cells involved in a pro-tumor immune response that are contained in the removed lymphoid tissue.

Abstract: Provided is a cell culture apparatus for culturing cells, and optionally, for performing magnetic separation of cells desired to be cultured. The cell culture apparatus preferably comprises a frame; at least one gas permeable membrane securedly sealed in a leak-proof sealing to a frame and an opposing surface comprising a rigid surface in forming a culture chamber therebetween; and at least one resealable aperture to allow substances to be introduced into, or withdrawn from, the culture chamber.

Abstract: Provided for heating cultured cells is a controlled heater device comprising a cell culture apparatus, a transparent heater in thermal contact with the cell culture apparatus, and may further comprise one or more of a transparent filter provided to prevent a predetermined spectral range of light from passing through the transparent filter, and a power source for controlling the amount of heat generated by the transparent heater of the controlled heater device.

Abstract: Compositions are provided for detecting Tn antigen expressing-shed tumor antigen. The compositions include peptides having binding specificity for the Tn antigen; and a detector molecule comprising a peptide, according to the present invention, which is bound to a detectable label. The compositions may be used in a method to detect lymphoid tissues containing shed tumor antigen expressing the Tn antigen.

Abstract: A method is provided for immune corrective surgical removal of lymphoid tissues containing shed tumor antigen in an individual. A detector molecule, with binding specificity for shed tumor antigen, is administered to an individual, and detected in the individual is the presence of lymphoid tissue which contains shed tumor antigen, as detected by the signal emitted by the detector molecule bound to shed tumor antigen. The lymphoid tissue, detected as containing shed tumor antigen, may then be surgically removed, thereby removing B cells, shed tumor antigen, and follicular dendritic cells involved in a pro-tumor immune response that are contained in the removed lymphoid tissue.

Abstract: Provided are a functionalized, encapsulated fluorescent nanocrystal comprising a liposome having encapsulated therein one or more fluorescent nanocrystals; use of the functionalized, encapsulated fluorescent nanocrystals in detection systems; and a method of producing functionalized, encapsulated fluorescent nanocrystals. A method of using the functionalized encapsulated fluorescent nanocrystals having affinity molecule bound thereto comprises contacting the functionalized encapsulated fluorescent nanocrystals with a sample so that complexes are formed between the functionalized encapsulated fluorescent nanocrystals and substrate for which the affinity molecule has binding specificity, if the substrate is present; exposing the complexes in the detection system to an excitation light source, and detecting a fluorescence peak emitted from the complexes, if present.

Abstract: Provided is a method of fluorescence imaging of living tissue using functionalized nanocrystals. The method comprises contacting an effective amount of functionalized nanocrystals with the living tissue; exposing the tissue to a spectrum of light suitable for exciting functionalized nanocrystals, present in fluorescently labeled tissue, to emit a emission spectrum comprising a fluorescence peak; and detecting any fluorescence peak emitted by the tissue exposed to the excitation spectrum of light, and obtaining a fluorescence image of the tissue. Also provided is a composition comprising a functionalized nanocrystal which is bound to a substrate in a living tissue.

Abstract: Provided are methods for purifying functionalized fluorescent nanocrystals having affinity ligand operably bound thereto by using a solid support matrix in a reactor through which solutions are circulated, and by using an immobilized solid phase formed by operably binding functionalized fluorescent nanocrystals to the solid support matrix. Also provided are purified functionalized fluorescent nanocrystals having affinity ligand operably bound thereto which are purified using the methods according to the present invention to be substantially free of free affinity ligand.

Abstract: Provided are a fluorescent microsphere comprised of a plurality of fluorescent nanocrystals operably bound to a polymeric microsphere, and a method of producing the fluorescent microspheres which comprises contacting the polymeric microsphere with a plurality of fluorescent nanocrystals under suitable conditions in which the fluorescent naocrystals become operably bound to the polymeric microsphere. Also provided is a method of using the fluorescent microspheres capable of determining the presence or absence of a predetermined number of analytes in a sample by contacting the sample with the fluorescent microspheres, and detecting the fluorescence signal pattern of excited fluorescent microspheres bound to one or more analytes of the predetermined number of analytes, if present in the sample.