Extended Data Fig. 1: Full optical diagram of Image-seq set-up.
From: Image-seq: spatially resolved single-cell sequencing guided by in situ and in vivo imaging
Radiation at 775 nm is generated by frequency doubling the 1550 nm output from a fiber laser. Radiation at 920-980 nm is generated by soliton self-frequency shift of 1550 nm radiation to 1840–1960nm in a LMA photonic crystal fiber, which is subsequently frequency doubled to obtain the imaging wavelength. A scanning GM (y scan) and scanning PM (x scan) are conjugated to one another, along with the back aperture of the OL so that a scan along the x and y dimensions is achieved in the image plane. Multiphoton emission wavelengths are collected with a combination of DCs and PMTs to generate an image. A confocal reflectance channel that utilizes an APD and 20µm-size pinhole can also be used to generate images. 𝛌/2: 𝛌/2-plate. PBS: polarization beam splitter. BiBO: BiBO3 - Bismuth orthoborate crystal. LMA: large mode-area. APD: avalanche photodiode. ph: pinhole. DC: dichroic mirror. GM: galvo mirror. PM: polygonal mirror. PMT: photomultiplier tube. MP: micropipette. OL: objective lens.
