Reference grid assembly for use in expansion microscopy
AssigneeUniversiteit Utrecht Holding B.V.
InventorsHugo Geert Jan DAMSTRA, Josiah Benjamin PASSMORE, Wilco NIJENHUIS, Lukas KAPITEIN
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Abstract
The present invention relates to a reference grid assembly for use in expansion microscopy (ExM) of a biological sample providing a scalable and expandible calibration marker for ExM. The present invention further relates to methods for analyzing a biological sample by expansion microscopy using the reference grid assembly and the use of the reference grid assembly in expansion microscopy of a biological sample.
Claims23
A reference grid assembly for use in expansion microscopy (ExM) of a biological sample, wherein the reference grid assembly comprises an expandible hydrogel or a substrate suitable for cell culture, wherein said hydrogel or substrate comprises a micropattern comprising micropatterned photoreactive or photoactivatable organic molecules or peptides providing a scalable and expandible calibration marker for ExM.
(canceled) (ExM)
(canceled)
Method according to claim 21, wherein the method further comprises the step of f) or e), respectively, quantitative interpretation of the expanded biological sample by determination of the expansion factor and deformation mapping using the reference grid.
The method according to claim 21, wherein the method furthermore comprises the step of contacting the micropatterned photoreactive or photoactivatable proteins with fluorescent antibodies providing further signal amplification of the micropatterned photoreactive or photoactivatable proteins.
The method according to claim 21, wherein micropatterning is done using deep UV or by laser, for example protein photolithography with deep-UV.
The method according to claim 22, wherein the culturing substrate comprised of the reference grid is sterilized before application of the biological sample on said culturing substrate.
The method according to claim 21, wherein the substrate is a coverslip, preferably a glass, polyacrylamide or polydimethylsiloxane coverslip.
The method according to claim 21-18, wherein said substrate is furthermore provided with a top coat, wherein the topcoat is comprised of poly-L-lysine and/or laminin.
(canceled)
A method for analyzing a biological sample by expansion microscopy (ExM) comprising:
- a) incorporating a biological sample in a reference grid assembly, the reference grid assembly comprising an expandible hydrogel or a substrate suitable for cell culture, wherein said hydrogel or substrate comprises a micropattern comprising micropatterned photoreactive or photoactivatable organic molecules or peptides providing a scalable and expandible calibration marker for ExM,
- b) expansion of said hydrogel including the biological sample and the reference grid by addition of an aqueous solution to the polymerized hydrogel,
- c) observing the expanded biological sample under a microscope.
Dependencies are inferred from the claim text.