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

Other

Beecher Instruments

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 Government of the United States of America, As Represented by the

Stephen B Leighton

James L Olds

John A Tenney

Michael Jay Brownstein

Norman E Rosenthal

Olli-P Kallioniemi

Juha Kononen

George Steven Bova

G Maret Maxwell

Alan M Kuzirian

Burt Chikadel

Allen Markowitz

Genesio Murano

William H Boyd

Thomas A Wehr

An array-based technology facilitates rapid correlated gene copy number and expression profiling of very large numbers of human tumors. Hundreds of cylindrical tissue biopsies (diameter 0.6 mm) from morphologically representative regions of individual tumors can be arrayed in a single paraffin block. Consecutive sections from such arrays provide targets for parallel in situ visualization and quantitation of DNA, RNA or protein targets. For example, amplifications of six loci (mybL2, erbB2, Cyclin-D1, myc, 17q23 and 20q13) were rapidly determined by fluorescence in situ hybridization from 372 ethanol-fixed breast cancers. Stratification of tumors by estrogen receptor and p53 expression data revealed distinct patterns of gene amplification in the various subgroups of breast cancer that may have prognostic utility. The tissue array technology is useful in the rapid molecular profiling of hundreds of normal and pathological tissue specimens or cultured cells.

Tumor tissue microarrays for rapid molecular profiling

Arrays of biological tissue can be created by removing cores from regions of interest in a series of donor blocks of embedded tissues. The cores removed are placed in a regular array in a recipient block. This is typically done with two different punches, one for obtaining the cores of interest and the other for creating the receiving holes in the recipient block. The present invention comprises such a system including a single z axis, with a mechanism for automatically changing two or more punches in and out of a holder on the z axis.

Punch-changing tissue array instrument

A method and device for storing and dispensing specified subsets from a collection of tens, hundreds or thousands of different reagents to specified locations within microtitre trays or microarray substrates or the like. The reagents of interest are stored in racks of syringe-like dispensers. These dispensers and/or substrates and/or actuators are controllably moved by automated means to a dispensing position where a single action, vibration and weighing system transfers a controlled quantity to a specified location in a microtitre tray or the like. Deficits of the prior art, such as evaporation, cross-contamination, oxidation, waste of reagents and lab-ware, and human operator non-reliability are overcome.

Method and apparatus for storing and dispensing reagents

Arrays of biological tissue can be created by removing cores from regions of interest in a series of donor blocks of embedded tissues. The cores removed are placed in a regular array in a recipient block. This is typically done with two different punches, one for obtaining the cores of interest and the other for creating the receiving holes in the recipient block. The present invention comprises such a system including two separate z axes, one for each punch. Each punch has its own stylet and the axis of each punch is parallel to the axis of its drive.

Double z-drive tissue array instrument

A method and device are provided for simultaneously creating a plurality of identical micro-arrays of biological samples. The invention utilizes a plurality of substrates, each of which having a top side, a bottom side, and a pattern of through-holes. Each through-hole has a wider upper cross-section, a narrower lower cross-section, and a step or plateau formed in the transition area. When the substrates are stacked, through-holes are in registry and form tunnels extending through the stack of substrates. Reagents of interest are caused to flow through the tunnels and deposit on the step or plateau area. A barrier layer may be provided to prevent leak-through between neighboring holes. After the desired reagents have been deposited, the substrates are separated. In this manner a series of micro-arrays, each capable of containing hundreds or thousands of biological samples such as cDNA fragments, is formed simultaneously.

Multiple micro-arrays

A simple, robust and precise instrument for constructing tissue arrays. The instrument includes multiple punches mounted on a punch platform, the punch platform displaceable between precisely defined positions. Mechanical detentes or stops are provided which mechanically arrest the movement of the punch platform in the precisely defined positions. This arrangement greatly saves time and improves accuracy over use of conventional precision linear positioning means. By the simple step of moving the punch platform from a first position to a second position, either punch can be quickly brought into operating position (and the other moved into a non-interfering position) by either manually or by automatic means, making it possible to quickly alternate from one punch to the other.

Instrument for constructing tissue arrays

A two- and three-dimensional autoradiographical imaging system is provided which includes a charge coupled device for detecting the emission of radioactively labeled substances from materials such as tissue samples, brains of humans or animals, or substances used in electrophoresis applications. In a first aspect, a radioactively labeled substance is included in a tissue sample. The tissue sample is sequentially imaged by a charge coupled device and a sectioning tool such as a microtome to produce a plurality of two-dimensional images. A three-dimensional image of the tissue sample is generated by further processing of the plurality of two-dimensional images derived from the charge coupled device. In a further aspect of the invention, a charge coupled device is utilized to provide realtime imaging of metabolic or physiological parameters involved in brain activity. A charge coupled device is positioned adjacent a portion of brain tissue desired to be examined, the brain tissue having a radioactively labeled substance therein for detection by the charge coupled device. A two-dimensional realtime image is produced using the charge coupled device for use in clinical or behavioral studies. In another aspect of the invention, a charge coupled device is utilized in electrophoresis applications to monitor radioactively labeled or tagged substances during electrophoresis. In this application, use of the charge coupled device permits small sample volumes of material to be used during electrophoresis, as well as automatic end point detection.

Method and apparatus for producing an image of a substance using a

A two- and three-dimensional autoradiographical imaging system is provided which includes a charge coupled device for detecting the emission of radioactively labeled substances from materials such as tissue samples, brains of humans or animals, or substances used in electrophoresis applications. In a first aspect, a radioactively labeled substance is included in a tissue sample. The tissue sample is sequentially imaged by a charge coupled device and a sectioning tool such as a microtome to produce a plurality of two-dimensional images. A three-dimensional image of the tissue sample is generated by further processing of the plurality of two-dimensional images derived from the charge coupled device. In a further aspect of the invention, a charge coupled device is utilized to provide realtime imaging of metabolic or physiological parameters involved in brain activity. A charge coupled device is positioned adjacent a portion of brain tissue desired to be examined, the brain tissue having a radioactively labeled substance therein for detection by the charge coupled device. A two-dimensional realtime image is produced using the charge coupled device for use in clinical or behavioral studies.

Two- and three-dimensional autoradiographic imaging utilizing charge

A microtome for cutting thin sample sections utilizes an electrical contact or sole plate for guiding the section cutting knife during sectioning. The sole plate floats across the surface of the sample thereby ensuring that uniformly thin sections are cut regardless of movement of the sample or expansion or contraction thereof. An electrical contact may be used to detect and reference contact between the sample surface and the section cutting knife. After contact between the sample surface and section cutting knife is referenced, the relative position between the sample surface and section cutting knife may be adjusted to achieve a desired section thinness.

Microtome with micro-plane and electric contact reference

An injection device injects small amounts of injectate into each of a plurality of cells. A first plate, which may be made from a silicon chip, includes a large number of evenly spaced cell wells, in a regular array. Each well includes a through hole in the chip. A top plate, also formed from a silicon chip, includes a plurality of injection needles, arranged in a regular array corresponding to the position of the cell wells. A suspension of cells is washed over the first plate and individual cells are retained by the individual wells. A vacuum may be applied to the cell wells through a manifold beneath the first plate, thus to retain the cells in place. Excess cells and suspension are removed by washing the plate. The second plate, including the injection needles, is positioned on the first plate so that the needles pierce the plurality of cells retained in the wells, thus simultaneously introducing the injectate into a large number of individual cells.

Array-type multiple cell injector

A prosthetic urethral sphincter valve with an integral spring valve member which comprises an elastic valve element having an upper portion which defines a central fluid passage and a lower diaphragm portion which includes a rolling diaphragm. The prosthetic urethral sphincter valve is placed totally within a patient's urethra. The lower diaphragm portion of the elastic valve element includes a tapered wall structure which provides for a spring action which demonstrates a non-linear force curve. The central fluid passage assumes a kinked or closed position, or a straighten or open position depending upon the position of the rolling diaphragm. Applied bladder pressure effects the position of the rolling diaphragm and thus the opening and closing of the central fluid passage.

Intra-urethral valve with integral spring

A device for delivering high intensity light to a patient's eyes for treating seasonal affective disorder and the like uses a point source of light such as a high intensity halogen or other incandescent bulb, and directs a large fraction of the light from the bulb directly into the patient's eyes without focusing the light in such a way as to cause damage to the eye or discomfort to the patient. This is accomplished by the use of a positive lens which focuses the light from the high intensity bulb directly in front of the patient's eyes. The light appears to the patient to be coming from an area much larger than the actual point source, and hence is more comfortable for the patient. The patient is assured of receiving a significant dosage of light no matter which way he is directing his gaze.

Portable light delivery system

A device for measuring the handedness, left and right, of human ambulatory turning behavior includes a rugged and compact sensing means in the form a Hall-effect compass or flux gate compass, as well as computation and storage and read-out elements for the total number of turns, half turns and quarter turns made by a patient wearing the device.

Magnetic personal turning monitor

A sensing device for obtaining optical density or light scattering measurements or other, in a turbulent liquid comprising a light source and a facing sensor located within a housing. The housing is provided with orifices through which liquid flows into the housing for measurement. The liquid entering the housing is slowed by passage through the orifice, and the bubbles of the liquid rise to the upper region of the housing, out of the measurement region, e.g. out of the line of the sight between the light source and the sensor.

Method of sensing fluid properties independent of bubble concentrations

A prosthetic urethral sphincter valve that is placed totally within a patient's urethra, the valve including a collapsible flexible thin-walled annular bag member secured in a rigid casing with flexible retaining petals at its top end receivable in the patient's bladder. An upstanding annular flexible thin-walled diaphragm is provided over an inturned top flange on the casing, the flange having small flow apertures forming damping ports communicating with the flexible bag member. The bottom end of the bag member is engaged with an annular guide urged upwardly by a coiled spring bearing on an inturned bottom flange of the casing. The working space between the bag member and the diaphragm is filled with viscous grease. The bag member has a central tubular passage which is normally occluded or kinked by the upward biasing force of the coiled spring. When the patient exerts sustained bladder pressure, the flexible bag member is distended downwardly, causing the central tubular passage to elongate and thereby open up, enabling it to pass urine. After urination, the bladder pressure is released, allowing the spring to return the bag member to its collapsed state and restore the occlusion of its central tubular passage, providing positive shut-off of the valve.

Growing community of inventors

Maplewood, NJ, United States of America

Stephen B Leighton