Correlated Microscope


Figure 1: Eukaryotes versus prokaryotes. (a) Eukaryotic cells are typically >10 μm in diameter and possess membrane-bound organelles such as a nucleus and mitochondria. (b) Example of a single-celled eukaryote, Euglena gracilis, in brightfield. (c) Fluorescence image of Euglena gracilis. (d) Algae, such as these Acrochae- tium, often form complex assemblies of cells. (e) Prokaryotes—bacteria and archaea—are typically 1–2 μm in diameter and have no membrane-bound organelles. The DNA is located in a “nucleoid” region, but no nuclear membrane is present. (f) The highest resolution light micrographs reveal few cellular features. This image shows the gut bacterium Escherichia coli in phase contrast. (g) Dyes targeted to membranes or nucleic acids can show some cellular features of prokaryotes. This image shows the soil bacterium Bacillus subtilis labeled with the membrane dye FM1-43.


Depending on instrument goals, this can be partially mitigated by increasing the detector array size and appropriate lenslet array selection. However, the capability of synchronous volu- metric imaging could offer game-changing prospects for many applications, particularly when combined with a DHM that provides high resolution in both amplitude and phase imaging.


20 Here we present a design for a combined DHM/FLFM


instrument and quantify its performance using test samples. Te trade-offs in volume versus resolution are discussed and future improvements suggested. In a second paper to be pub- lished in Microscopy Today, we will present data collected from cultured bacterial and in situ environmental microorganisms


www.microscopy-today.com • 2020 May


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