Embedded Files
Protocol for Quantifying Fos Immunohistochemistry Results using ImageJ
Stitching images together into a larger image
Staining intensity quantification protocol
Cryotome Instructions
Cryotome Instructions
(a.k.a. freezing sliding/sledge microtome)
Document: https://docs.google.com/document/d/1n1wyyahoLdlC-CDyxgFad66iwwvrhICw5Hn7oqj9Gt0/edit?usp=sharing
Associated video: Will update later!
Vibratome tissue sectioning protocol
Vibratome tissue sectioning protocol
Vibratome Troubleshooting:
Getting Better Quality Sections
Vibratome Troubleshooting:
Getting Better Quality Sections
Recipe for PVA coverslipping medium
Recipe for PVA coverslipping medium
Have you used Permount or DPX as coverslipping medium for your slides, but then you have to deal with those substances being hazardous? Or potentially incompatible with certain fluorescent reporters? I suggest using polyvinyl alcohol medium, which you can make in your lab.
Recipe document: https://docs.google.com/document/d/1H-UniGEAO3Aq9YT7CR4cY7NMiLFIZdBTE2IHr0FSy80/edit?usp=sharing
Tyramide conjugate synthesis
Tyramide conjugate synthesis
Tyramide signal amplification is an approach that allows for fluorescent dyes to be continually deposited around a target as long as the reaction progresses. This technique is very helpful for amplifying dim immunohistochemistry or in situ hybridization signals. Ready-to-use tyramide conjugates are costly from vendors, so why not make your own in lab? You can make far larger amounts for a fraction of the potential cost.
Original article by Hopman et al. (1998) explaining tyramide conjugate synthesis: https://doi.org/10.1177%2F002215549804600611
(Note some typos: In the biotin recipe, the Stock B Tyramine volume is ten times too high - should be 29 uL, not 289 uL. Also, the mass of dye ester used should be mg not ug.)
My protocol: https://docs.google.com/document/d/1VrDpUDp-hXczqOSgKXquACqEIBOMTTgqupseyY6CSeM/edit?usp=sharing
Multi-label IHC protocol using multiple rounds of tyramide signal amplification
Multi-label IHC protocol using multiple rounds of tyramide signal amplification
Did you know that multiple rounds of tyramide signal amplification can be applied to the same tissue, using different dye colors, to label distinct targets without accidental cross-labeling? Here's the info on how and why to do it.
Video on concept of how it works and why I use it: https://youtu.be/u_JswJUwzts
Demo Video: https://youtu.be/aKsMS5cwTvs
Protocol used in demo video: https://docs.google.com/document/d/1djb6CZKEoGbauZpYMAwdukBkQnbr3H8S8m4ZaOPBgCs/edit?usp=sharing
Schematic files for 3D printing
Schematic files for 3D printing
Netwell inserts + basins
Netwell inserts + basins
I replicated the small netwells used in my videos in TinkerCAD online. You'll still need to purchase a mesh (vinyl or some other plastic; check McMaster-Carr or a similar industrial supply website) to serve as the net and glue it on, but a 3D printer will make the rest.
6-well insert without legs: https://drive.google.com/file/d/1fK8YsjlMDidNOkAhgc61pXsRjMgdvD8L/view?usp=sharing
6-well insert with legs: https://drive.google.com/file/d/1hDHAEqGx1SrzkAtWC5injG1xrj1DXM68/view?usp=sharing
Basin for 6-well insert: https://drive.google.com/file/d/15V4LqvDtnnwLOE2nZQJJ8sgu3e_Pt9K6/view?usp=sharing
24-well insert without legs: https://drive.google.com/file/d/1V7yuVGwm9CC9JvZffFtCYGMvibbqhBcV/view?usp=sharing
Basin for 24-well insert: https://drive.google.com/file/d/15NSpzdh9pKvtAx0FxLgzJmlfKy-wBrXN/view?usp=sharing
Vibratome parts
Vibratome parts
Folder containing designs for various parts that replace the Leica vibratome blade holder and the basin: https://drive.google.com/drive/folders/1mYDHy5GGGqrEse5euZQKfBDFhAkJyG7v?usp=drive_link
See the notes document (in this folder) to find additional parts - screws and tap drills - you would need to purchase.
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