Eye of the Beholder

The human eye is estimated to have a resolution of 576 megapixels. The camera attached to my laboratory’s microscope has 8. That’s the same amount of pixels an iPhone camera collects, and while it may be true that one could capture breathtaking images that convey everything a viewer may wish to know in one shot… those images will never hold a candle to what can be seen with the human eye.

What happens then, when the human eye is removed from the equation? Many microscopes these days are being built without eyepieces, and instead are manipulated and viewed entirely through (often proprietary) software. This not only cuts down costs for both the manufacturer and the customer (optics are expensive to fashion and align), but also allows for fun environmental parameters such as live cell imaging with temperature and carbon dioxide level controls. And don’t get me wrong, these machines can take beautiful images- check out the picture below if you don’t believe me.

MAX_Composite Vimentin 10x

Dismorphic vasculature in the mouse kidney.

However, with immunofluorescence in particular, a common problem plaguing interpretation of data is autofluorescence, or background fluorescence. When you are looking through the eyepiece of a standard confocal microscope, your eye usually does a pretty good job at discerning the true signal from the false autofluorescence. This in turn allows you to adjust the camera’s settings such that you acquire a biologically accurate image.

But with these software controlled microscopes, using a camera as an eyepiece means that you cannot easily detect the nuances in emission from your fluorophore. Already I’ve begun to see images creeping into the literature that are overexposed, full of background, and are generally of poor quality and doing little to highlight biologically significant features. That is why I am asking you, dear reader, whether you be a student, post-doc, PI, or anywhere in-between, to always opt for optics. That fanciest software in the world is no match to the power of the human eye.