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 Secretary of

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

Other

The United States of America

Seth R Goldstein

Paul D Smith

Robert F Bonner

John I Peterson

Thomas J Pohida

Lance A Liotta

David B Krizman

Michael R Emmert-Buck

W Marston Linehan

Rodrigo F Chuaqui

Jeffry M Trent

Robert L Jones

Robert Jones

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 method of microdissection 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 extraction is achieved by contacting the tissue sample with a transfer surface that can be selectively activated so that regions thereof adhere to the zone of cells of interest to be extracted. The transfer surface includes a selectively activatable adhesive layer which provides, for example, chemical or electrostatic adherence to the selected regions of the tissue sample. After the transfer surface is activated, the transfer surface and tissue sample are separated. During separation, the zone of cells of interest remains adhered to the transfer surface and is thus separated from the tissue sample, the zone of cells of interest may then be molecularly analyzed.

Isolation of cellular material under microscopic visualization

A machine for automatically tying a four-in-hand necktie knot in a necktie, includes ( ) a horizontal rotatable cylinder; ( ) a hooking mechanism capable of pulling a left hand short segment of the necktie through a loop of a right hand long segment of the necktie hanging from the rotatable cylinder; ( ) a finger mechanism capable of laterally moving the right hand long segment along the length of the rotatable cylinder; ( ) a whirler mechanism capable of flipping an end of the right hand long segment around the rotatable cylinder and up through a space between the right hand long segment and the left hand short segment and the necktie support, and ( ) an electronic and feedback control for operating various mechanisms in response to a sequence of voltage commands.

Machine that ties and unties a four-in-hand necktie knot

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

A confocal scanning laser microscope has no moving parts and displays real time video images. The image of the scanned laser spot on the specimen is focused on the photocathode of an image dissector tube having an electron image deflection that is synchronized and aligned with the laser scan. This permits the confocal pinhole to be scanned at video rates. Acousto-optic deflectors are responsive to the frequencies of respective scan control signals to effect horizontal and vertical laser beam deflection.

Confocal scanning laser microscope having no moving parts

Disclosed is a method for obtaining access to a relatively inaccessible region of a blood vessel for diagnostic or therapeutic purposes in which a primary catheter tube is inserted into the vascular system at an entry point remote from the relatively inaccessible region, the leading end of the tube is worked towards the inaccessible region in a conventional manner and then a secondary catheter tube contained within the primary tube is everted from the leading end of the primary tube to approach more closely to the required region. A catheter assembly for performing the method is also disclosed.

Method and apparatus for traversing blood vessels

A fiber optic pH probe suitable to be implanted in tissue for physiological studies is disclosed. The probe includes an ion permeable membrane envelope which encloses the ends of a pair of optical fibers. A pH sensitive dye indicator composition is present within the envelope. The probe operates on the concept of optically detecting the change in color of a pH sensitive dye.

Fiber optic pH probe

An electrically controlled magnetic fluid actuated device, which can be used in controlling fluid flow and which has particular utility in controlling this flow from a pressurized reservoir implanted in the body such as might be used for an artificial pancreas, sphincter for bladder control or other orthotic devices in the body of a human, has no freely moving parts and consists of a flow passageway having a portion the cross-section of which may be varied to control and to stop the flow of fluid, such as medicine, from a reservoir in the body to the point where this fluid is needed. A magnetic fluid which surrounds or contacts this portion of the passageway can, according to the presence or lack of a magnetic flux in its vicinity, occlude the passageway or allow it to open so as to control the fluid flow in the flow member.

Growing community of inventors

Bethesda, MD, United States of America

Seth R Goldstein