Figure 1 shows a schematic of the constructed confocal fluorescence microscope [Fig. 1(a)] and a photo of the current setup [Fig. 1(b)]. The upright confocal microscope has a 40 mW, 488 nm, continuous wave, externally doubled diode laser (Excelsior, Spectra-Physics, Santa Clara, CA) for fluorescence imaging. The laser beam passes through a spatial filter, which also serves as a beam expander, and reflects off of a dichroic mirror (z488rdc, Chroma Technology Corporation, Rockingham, VT). A custom-designed polygon scanner (Lincoln Laser, Phoenix, AZ) with a scan rate of 2.5 to 8.75 kHz serves as the line scan, and a galvanometer scanner (6220H M40B, Cambridge Technology, Lexington, MA) with a scan rate up to 15 frames per second (fps) provides the frame scan for image acquisition in traditional raster-scanning mode, producing “square” two-dimensional images. Two telescopes are used to image both mirrors onto the back aperture of the , 0.8-numerical aperture, 3.5 mm working distance water immersion microscope objective lens (MRD07420, Nikon). The sample is positioned on an XYZ-motorized stage (KT-LSM100A, Zaber Technologies Inc, Vancouver, British Columbia, Canada) with 0.05 µm resolution, 100 mm of travel in each axis, and maximum speed. Fluorescence is excited in the sample, collected by the objective, passed back through the telescope system, and descanned by the scanning mirrors. After passing through the dichroic mirror to the detection arm, the signal is focused onto the confocal detection pinhole and then refocused onto a photomultiplier tube (PMT) detector (H9433-03MOD, Hamamatsu, Bridgewater, NJ) with gain set to and frequency bandwidth of 10 MHz. A frame grabber (NI PCI-1410, National Instruments, Austin, TX) digitizes the PMT signal with a 29 MHz pixel clock and generates images. Both of the scanning mirrors and the stage are controlled by LabVIEW software (National Instruments), and the mirrors are driven by a data acquisition (DAQ) board (NI PCI-6251, National Instruments).