**SUPER IMPORTANT** After the dissection, clean off the counter with Mr. Clean's multipurpose cleaner, then warm tap water, and finally ethanol–wiping it down with paper towels in between steps. This prevents the accumulation of (hard to remove) organic material on the counter.
If you are the last one to use the counter, be sure to spray and wipe down the counter and tools with warm water and 70% EtOH.
Do NOT keep the saline bottles out of the fridge for longer than necessary. Please refill them when they are close to empty and wipe down the bottles when they look dirty.
Wipe off any saline you spill - salt builds up quickly! (on the counter and in the fridge)
When leaving preparations in the fridge, label them with Sharpie with your name and date. Do not leave partially dissected preparations in the fridge for >1 day.
Please replace empty paper towel boxes, glove, or kimwipe boxes, refill transfer pipet boxes, replace old sandpaper, etc. (Ask if you don't know where these stocks are located.)
The carcass freezer gets emptied by the technician or undergrads when necessary. Let someone know if it is getting full and needs to be emptied.
Please refer to the published paper on the STNS Dissection. Several resources are also below.
Watch the Cancer borealis STNS dissection video (below).
Additional Tips:
Gross Dissection
Holding the rongeurs so the convex side is facing away from you can help with breaking parts of the carapace off.
Fine Dissection
Adjust the magnification on your oculars and the distance between them to your liking.
Be aware of your posture when dissecting (keeping a straight back minimizes discomfort).
Pin down the stomach tight + flat against the black dish to ensure good optics.
Adjust light sources and change out saline as often as necessary.
Make sure to keep track of the left + right sides of the dissection - you will not be able to access desheathed STG cells if the prep is pinned upside down.
When removing the prep from the dish, you can start by separating the lower-end nerves from the tissue, then move upward to the ions.
Make sure to clean your microdissection tools after you finish the fine dissection. (You can use tap water and EtOH to keep them clean and prevent rusting.)
These tools are expensive, so protect them. You can learn how to sharpen them, but they can be hard to repair if bent/blades get notched.
To avoid dry spots, you can use sandpaper to sand down the Sylgard dish before conditioning it with excess stomach tissue. (If you plan to record intracellularly from the cells, make sure not to sand down the dish too much in the middle where the STG will be. This helps maintain good optics in later steps.)
You can cut pins ahead of time and store them in a small Sylgard dish for later use.
For step-by-step guides of the complete C. borealis dissection with pictures refer to the following guides.
Cancer borealis Gross Dissection Guide
Cancer borealis Fine Dissection Guide
Watch the Homarus americanus gross dissection and fine dissection videos. The gross and fine dissection guides are below.
Homarus americanus Gross Dissection Guide
Homarus americanus Fine Dissection Guide
**Please ask someone for a general introduction to these tools before using them for the first time!***
Use the sharpening block and surgical instrument oil to grind away parts of the tools that are notched, misaligned, blunt, etc.
As you do so, frequently use a kimwipe to wipe off the oil and examine the points.
The oil sometimes bends light in ways that make you think the tools have been completely fixed when that is not the case.
Do your best to grind sparingly - the more steel you grind away, the smaller your point of contact (for forceps), and then the tools don't work as well.
If you are trying to repair forceps that became bent at the tip, use round nose pliers to gently and carefully bend the tip(s) back into alignment and then sharpen them together.
Whetstone for sharpening dissection tools and a small bottle with surgical instrument oil.