Image – Page 11 – The Live Cell Imaging facility

Are there brighter versions of GFP or RFP out there?

Have you ever heard about Superfolded GFP? It is 50% brighter than GFP! And mScarlet is almost 6 times brighter than mRFP! How do I know? I look at this fantastic database called FPBase.

You can see which fluorescent protein is monomeric, sort them by excitation and emission or find which bleaches least or maturates fastest! Great tool! 🙂

Fluorophores are constantly being developed. If you make a new plasmid, make sure you check that the one your lab has been using for trillions of years is the very best one!

Great video tutorials for our Primo users and paper on microniches

Hello again

Here you can see very nice video tutorials on the Alveole website and this is a cool article by Viasnoff et al about making 3D microniches with 1 um resolution! And you can do this at the LCI facility!! 🙂

Silicon wells

If you want to image a large piece of tissue, it is sometimes difficult to get it to stay still in a dish while you are imaging it, especially if you have medium on top.

One nice way is to make a silicon well around it, fill the well with medium then add a coverslip on top. This allow you to keep your sample for a long time.

Twinsil by Picodent works nicely. 🙂

More about RNA labeling

Yet another chance to try RNA labelling: The FENO facility, here in Flemingsberg, has purchased a machine to multiplex RNA scope. They will present it on the 16th of October. 🙂

Here is the announcement.

Robust RNA labeling in cells and tissue

We got tipped by one of our users of a robust way to label and image RNAs in cells and tissue. Here is the paper and one can buy the kit from here. Apparently it works a charm! Let us know if it works for you or not. 🙂

A beginner’s guide to tissue clearing

Someone telling you where to start your clearing experiments! Sounds like a dream paper, doesn’t it? 😉

Here is the link.

Omnipaque: a pre-made Histodenz solution for clearing

Buying Histodenz powder from Sigma is rather expensive. Alternatively, you can buy Omnipaque from GE Healthcare. It is the same product but comes as a solution and costs around 2000 sek for 10 bottles of 350 mL!

A fixable membrane dye that doesn’t do flip flop!

One of the major pains with membrane dyes for live cells is that within 30min, all the vesicles inside the cells are also labelled. Another major pain is when you discover that your dye is not fixable…

The CellBright dyes label membrane is apparently fixable, even with methanol, and doesn’t do the flip flop thing! Potentially extremely useful for image analysis!

Please leave a comment to let us know how it went for you! 🙂

Clearing and expansion microscopy course at Scilife in September!

Expansion microscopy and clearing are fantastic tools for anyone who images thick (> 1 cell diameter) fixed samples. We are now lucky to have access to state of the art talks and hands-on workshops at Scilife in September. Register early not to be disappointed!

Please apply to Hans (first day) and David (second day) directly!

Seminar by Anne Carpenter, the ‘mother’ of CellProfiler

If you are looking for an inspiring talk about a super freeware for image analysis, go to Scilife in Uppsala on the 8th of June and listen to Anne Carpenter from the Broad Institute where CellProfiler was born! Check out the details here.