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The 647 nm line of a water-cooled Kr+ laser (Innova 302, Coherent, Santa Clara, CA) at 400 mW was injected into an integrated Olympus microscope and focused to a spot size of approximately 2 μm (100× or 20× objective).
Activation (405 nm/532 nm) and excitation (561 nm/641nm) lasers for PALM/dSTORM were combined in an external platform; the combined beam was expanded and re-collimated with a beam expander, directed through free air into the microscope and focused in the rear plane of a 100× objective (N.A. 1.49) using an achromatic converging doublet-lens with a focal length of 500 mm.
The laser was then directed into a Nikon TE-2000 microscope and focused into the sample by a 0.8 NA, 60x air objective (Nikon).
The laser is then, coupled into the objective lens via the side port of the microscope, and focused onto a diffraction-limited spot with the objective lens.
The laser was coupled to a spinning-disc inverted microscope and focused to axons for severing (Fig. 3A) or neuronal soma for ablation (Fig. 3C,D) in Tg[SILL mCherry] transgenics.
The diode laser was coupled with a liquid fiber of 200 µm diameter to the epifluorescence path of the microscope and focused on the slice with the ×40 objective.
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I was grateful to be able to put my scrubs back on, peer into the oculars of the microscope, and focus on the task at hand, to take a surgical approach to fix a problem.
Using a 5× lens on a Zeiss wide-field microscope and focusing on the middle of the sample, we image the local density of these particles, which displays heterogeneities as seen in figure 2E.
The laser light was then directed to a 60X microscope objective and focused onto a 2.1 µm silica suspended-core fiber, shown in the inset of Fig. 1.
When studying the dynamics using Fluorescence or Dark/Bright field microscope, accuracy and focusing problems exist due to the MF movement in all directions as well as their frequent coiling [ 35].
In this paper we present simple procedures to couple LED based illumination sources to a standard widefield Olympus microscope and focus on one imaging application that employs blue and infrared LEDs.
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