Method and system for imaging and analysis of a biological specimen
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsKarl A. Deisseroth, William E. Allen, Brian Hsueh, Li Ye
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Abstract
The present disclosure provides methods of preparing a biological specimen for imaging analysis, comprising fixing and clearing the biological specimen and subsequently analyzing the cleared biological specimen using microscopy. Also included are methods of quantifying cells, for example, active populations of cells in response to a stimulant. The present disclosure also provides devices for practicing the described methods. A flow-assisted clearing device provides rapid clearing of hydrogel-embedded biological specimens without the need of specialized equipment such as electrophoresis or perfusion devices.
Claims29
A method of preparing a biological specimen for imaging analysis, the method comprising:
- i) fixing the biological specimen with a plurality of hydrogel monomers to produce a hydrogel-fixed specimen;
- ii) clearing the hydrogel-fixed specimen using a non-electrophoretic flow-assisted clearing device to produce a cleared specimen, wherein the clearing is performed by:
- a) immersing the hydrogel-fixed specimen in a buffer in a sample chamber of the flow-assisted clearing device;
- b) flowing the buffer through the hydrogel-fixed specimen by using a buffer circulator, wherein buffer flow through the hydrogel-fixed specimen is unidirectional and sufficient to clear the hydrogel-fixed specimen, wherein the buffer flow direction through the hydrogel-fixed specimen is determined by the buffer circulator;
- and
- iii) contacting the cleared specimen with a refractive index matching solution prior to imaging analysis.
Dependencies are inferred from the claim text.