See Cells In A New Light With A DIY Fluorescence Microscope

Ever since a Dutch businessman peered into the microscopic world through his brass and glass contraption in the 1600s, microscopy has had a long, rich history of DIY innovation. This DIY fluorescence microscope is another step along that DIY path that might just open up a powerful imaging technique to amateur scientists and biohackers.

In fluorescence microscopy, cells are treated with various fluorescent dyes that can be excited with light at one wavelength and emit light at another. But as [Jonathan Bumstead] points out, fluorescence microscopes are generally priced out of the range of biohackers. His homebrew scope levels the playing field a bit. The trick is to use 3D-printed parts to kit out commonly available digital cameras – a USB microscope, a DSLR, or even a smartphone camera. Excitation is provided by a ring of Nexopixel LEDs, while a movable rack holds a filter that blocks the excitation wavelength but allows the emission wavelength to pass through to the camera. He demonstrates the technique by staining some threads with fluorescent ink from a highlighter marker and placing them on a sheet of tissue paper; in conventional bright-field mode, the threads all but disappear into the background, but jump right out under fluorescence.

Ith’s true that the optics are not exactly lab quality, and the microscope is currently only set up to do reflectance imaging as opposed to the more typical transmissive mode where the light passes through the sample. That’s an easy fix, though, and reflectance mode is still useful. We’ve seen fluorescence microscopy get quite complex before, but this simple scope might be enough to get a biohacker started.

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Fluorescence Microscopy Meets DIY Fluid Management

Fluorescence microscopy is an optical technique that incorporates fluorescence or phosphorescence (as opposed to reflection and absorption) in order to study the properties of organic and inorganic substances. Not a stranger to bringing DIY techniques into the lab, [Philip] is using 3D printing resources to advance science and delight interns from labs everywhere.

In fluorescence microscopy, a huge limiting factor that decreases the amount of data that can be gleaned from a single sample is the number of targets that can be labeled with fluorescent tags. However, overlap in the spectral emissions of fluorophores limits the fluorophores that can be used side-by-side. This means that only around four targets can be labeled with fluorescent tags in a typical setup, with ten being the absolute maximum if careful spectral demixing is done. However, in a single sample, there might be a few hundred components. Clearly, we’re off by an order of magnitude (or more).

However, researchers are smart. One current solution is to label targets in a sequential manner with probe signal nullification steps in between. Ideally, probes are introduced in sequential without moving the sample off of the microscope. After imaging, the probes can be removed, allowing the number of labeled targets to be limited only to the number of rounds of probe replacement. And, with clever ‘barcoding’ schemes, the returns from each round can even scale exponentially, rather than linearly.

But, to accomplish this feat, a single sample must be processed through the labeling and stripping steps repeatedly. It’s not uncommon to do this by hand, consisting of many hours of exceptionally tedious work. That’s where [Philip] comes in. By using a 3D printer like Cartesian robot, [Philip] is automating the labeling and stripping steps resulting in happy interns and ultimately a more precise product. Rather than spending a few tens of thousands on a commercial machine, you can find all of [Philip’s] design files in his GitHub repo and make one for ~ $1k. Ready for more? We’ve got your back.

Video after the break.

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