The United States of America, As Represented by the Secretary, Department of Health and Human Services

The United States of America As Represented by the Department of Health

Thomas J Pohida

Robert F Bonner

Paul D Smith

Seth R Goldstein

Michael R Emmert-Buck

Rodrigo F Chuaqui

Michael Anthony Tangrea

John I Peterson

James B Mitchell

Walter S Friauf

Mark S Mirotznik

Sankaran Subramanian

Michael M Gottesman

Bradley Allen Roth

Rolf G Tschudin

Murali K Cherukuri

Nicole Y Morgan

Paul G Zablocky

John Bourg

Chandan Das

Philip G McQueen

John U Dennis

Ashley A Jaeger

Peter W Schuck

John W Kakareka

Hauying Zhao

Ghadi Salem

Randall H Pursley

George Patterson

The present disclosure provides various embodiments of a fluorescence scanning system having a sample holder with a sample suspended within that is rotated by a centrifuge such that the sample is illuminated at various angles by an excitation beam by operation of a galvanometer and such that the sample emits a fluorescence emissions that is detected through a narrow window of exposure defined along the travel path of rotation taken by the sample holder when rotated by the centrifuge. A stationary fluorescence detector is in operative communication with the sample holder along the narrow window of exposure for detecting the fluorescence emissions emitted by the sample from the sample holder while also separating the excitation beam from the fluorescence emissions.

Fluorescence scanning system for analytical ultracentrifugation

The System for Continuous Observation of Rodents in Home-cage Environment (SCORHE) was developed to quantify activity levels and behavior patterns for mice housed within a home cage that, optional, may be disposed in a commercial ventilated cage rack. The SCORHE home-cage in-rack design provides daytime and night-time monitoring with the stability and consistency of the home-cage environment. The dual-video camera custom hardware design makes efficient use of space, does not require home-cage modification, and is animal facility user-friendly. In various embodiments, a software application integrates the two video streams to extract several mouse activity measures.

Systems and methods of video monitoring for vivarium cages

The present disclosure is directed to embodiments of microstructured membranes, methods of fabricating microstructured membranes, bioreactors housing microstructured membranes, and methods of using bioreactors and microstructured membranes. In some embodiments, the present disclosure allows culturing of cellular tissues in an environment which more accurately resembles a native environment. In some more specific embodiments, the present disclosure allows culturing of tumor cells on a membrane having a microfabricated pattern which mimics a native vasculature system.

Microfabricated polymeric vessel mimetics

A method of removing a target from a biological sample which involves placing a transfer surface in contact with the biological sample, and then focally altering the transfer surface to allow selective separation of the target from the biological sample. In disclosed embodiments, the target is a cell or cellular component of a tissue section and the transfer surface is a film that can be focally altered to adhere the target to the transfer surface. Subsequent separation of the film from the tissue section selectively removes the adhered target from the tissue section. The transfer surface is activated from within the target to adhere the target to the transfer surface, for example by heating the target to adhere it to a thermoplastic transfer surface. Such in situ activation can be achieved by exposing the biological sample to an immunoreagent that specifically binds to the target (or a component of the target). The immunoreagent can alter the transfer surface directly (for example with a heat generating enzyme carried by the immunoreagent), or indirectly (for example by changing a characteristic of the target). In some embodiments, the immunoreagent deposits a precipitate in the target that increases its light absorption relative to surrounding tissue, such that the biological specimen can be exposed to light to selectively heat the target. Alternatively, the immunoreagent is an immunofluorescent agent that carries a fluorophore that absorbs light and emits heat.

Target activated microtransfer

A device for performing target activated transfer that includes a mounting surface for mounting a tissue sample; and a light source positioned to substantially uniformly irradiate both stained and unstained regions of the tissue sample with light energy that activates the reagent to selectively adhere the stained regions to a transfer surface. Also described is an automated system for transferring tissue from a tissue sample to a transfer substrate. The system includes means for holding a tissue section that includes targets specifically stained with an absorptive stain thereby resulting in a stained tissue surface, and a flexible transfer film that includes a lower thermoplastic layer in sufficient thermal contact with the stained tissue surface; an irradiating assembly configured to provide a predetermined uniform light dose to the entire tissue section; and means for applying a constant pressure to the transfer film during irradiation.

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. In its preferred configuration it uses the inertial confinement of the surrounding unmelted thermoplastic polymer (and the overlying rigid substrate) to force expansion of the melted polymer into the underlying tissue target. Utilizing the short pulse protocol, the targeted and extracted material can have a diameter equal to or smaller than the exciting beam.

Method of laser capture microdissection from a sample utilizing short pulse length

A tissue sample is conventionally visualized in a microscope. A selectively activated convex surface is provided, preferably at the distal end of a rod. This selectively activated convex surface when activated, typically with a laser through an optic light path in the microscope, provides the activated region with adhesive properties. At least one portion of the tissue sample which is to be extracted is identified. This identified portion is contacted with a portion of the selectively activated convex surface on the end of the rod. When the convex surface is activated, typically by exposure to laser light in the footprint of the desired sample, an adhesive transfer surface on the selectively activated convex surface is provided which adheres to the desired cells in the footprint of the desired sample. Thereafter, the adhesive transfer surface is separated from the remainder of the tissue sample while maintaining adhesion with the desired cells. Thus the desired portion of the tissue sample is extracted. The disclosed selectively activated convex surface is preferably utilized to collect desired tissue samples at more than one location on the same slide or from different slides. The collected tissue samples can thereafter be inspected if desired, as collected on the convex surface, and then liberated&mdash;as by dissolving the proteins of the samples. This can effectively concentrate rarely occurring cells in order to obtain enough pure material for analysis. A rod having a convex surface with the selectively activated material is set forth as a staple for use with the apparatus and process. Preferred shapes for the convex surface are disclosed as well as a method for coating rods with a resultant rod article.

Convex geometry adhesive film system for laser capture microdissection

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.

Non-contact laser capture microdissection

A method and apparatus of gathering by LCM identified cellular material from randorn locations on a tissue sample to designated locations on a transporting substrate enables convenient further processing. A transporting substrate has an identified mapped location for receiving identified cellular material. At least a segment of a selectively activatable coating is placed on the side of the transporting substrate in apposition to the tissue sample at the mapped location. The transporting substrate and sample are relatively moved to place the selectively activated coating at the mapped location in apposition to identified cellular material of the tissue sample which is to be extracted. Thereafter, the selectively activatable coating is activated and impressed or impressed and activated to form an adhesive region on the transporting substrate for adhering to the identified cellular material. Upon removal of the transporting substrate from the tissue sample, identified cellular material adheres to the transporting substrate at the mapped location.

Mechanical handling systems for laser capture microdissection

Precision laser capture microdissection utilizing short pulse length

A process of microdissection where a tissue sample is conventionally visualized in a microscope. A selectively activatable convex surface is provided, preferably on the periphery at the distal end of a rod. This selectively activatable convex surface when locally activated, typically with a laser through an optic light path in the microscope, provides the activated region with adhesive properties. The tissue sample has at least one portion, which is to be extracted is identified. This identified portion is contacted with a portion of the selectively activatable convex surface on the periphery of the rod. When the convex surface is locally activated, typically by exposure to laser light in the footprint of the desired portion, an adhesive transfer surface on the selectively activatable convex surface is activated which adheres to the desired cells in the footprint of the desired portion. Thereafter, the adhesive transfer surface is separated from the remainder of the tissue sample while maintaining adhesion with the portion of the sample. Thus the desired portion of the tissue sample is extracted. The disclosed selectively activatable convex surface is preferably utilized to collect desired tissue samples at more than one location on the same slide or from different slides. A rod having a convex surface with the selectively activatable material is set forth as a staple for use with the apparatus and process. Preferred shapes for the convex surface are disclosed as well as a method for coating rods.

Method utilizing convex geometry for laser capture microdissection

An EPR imager and spectrometer includes pulse generating system for generating broadband pulses having an RF carrier frequency that is not highly absorbed by biological samples. The pulse generating system includes up and down chirp convertors for frequency modulating a carrier frequency pulse and compressing the frequency modulated pulse to form a broadband excitation pulse of high energy. Such a machine could form the basis of a clinical imaging device capable of high sensitivity to free radical species in human patients.

Growing community of inventors

Monrovia, MD, United States of America

Thomas J Pohida