Tag Archives: Fluorescence

Divide and conquer

One of the more annoying things about fluorescence imaging is that it’s a bit like trying to describe our location in the universe. There’s no absolute point of reference, the values are rather arbitrary and you rely heavily on relative measures (like being 1AU from our local star).

This post will demonstrate some of the problems with quantifying basic fluorescent images and use a case study to show how ratiometric imaging (among other things) can be used to solve them.

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Know thy Image Part 2: Image Histograms

In the last post, we looked at the following image to decide if an intensity of 2000 at the tip of the arrowhead was bright or dim. Without knowing the bit depth of the image it’s impossible to tell.

Bright or not bright?

We posed that if this was a 12bit image, that’s fairly bright (2000/4096) but if it’s a 16bit image, then it’s very dim (2000/65536). But why would these two conditions look the same? The answer lies in the histograms and transfer functions…

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Know thy Image Part 1: Bit Depths

In the list of things that everyone who uses a microscope should know, this has to be near the top and yet I find a surprising number of people are either never taught it or don’t fully grasp the idea of Bit Depth. In this post and Part 2, we will deal with (almost) everything you need to know about bit depth, dynamic range and image histograms.

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