FISH hybridization suspension was applied onto slide with 10 µL mounting media (including DAPI). Then the slide was subjected to the Axio Imager Z2 fluorescence microscope (Zeiss, Germany) for scanning. Each sample slide was scanned automatically using Metafer 5 software. This software enables 4-channel fluorescent images of multiple fields covering the full area of sample for fluorescence detection of cells. 10x magnification (Zeiss Plan-Apochromat 10x/0.45 M27) was used to scan the sample slide and exposure times were auto-selected at 0.04 s for DAPI, 0.44 s for CEP8, 0.8 s for CD45 and fixed at 0.6 s for EpCAM, CK18, PD-L1 and Vimentin. The captured images were screened manually and CTCs are identified as CD45−, CD31−, DAPI+, tumor biomarker(s)+/- with aneuploid chromosome 8. All sample slides were reviewed by the same investigator.
Plan apochromat 10x 0.45 m27
Manufactured by Zeiss
The Plan-Apochromat 10x/0.45 M27 is a high-performance objective lens developed by Zeiss. It is designed to provide high-quality imaging with minimal aberrations and distortions. The lens has a magnification of 10x and a numerical aperture of 0.45, making it suitable for a wide range of microscopy applications.
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Lab products found in correlation
Lsm 880, by Zeiss (2 mentions)
Axio imager z2, by Zeiss (1 mentions)
Fvmpe rs, by Olympus (1 mentions)
Plapon60x, by Olympus (1 mentions)
Uplsapo10x, by Olympus (1 mentions)
Plan apochromat 63x 1.4 oil dic m27, by Zeiss (1 mentions)
Ucplfln20x, by Olympus (1 mentions)
Fv1200mpe, by Olympus (1 mentions)
Calcofluor white, by Zeiss (1 mentions)
Fluorol yellow, by Zeiss (1 mentions)
Fluorol yellow, by Santa Cruz Biotechnology (1 mentions)
Plan apochromat 20x 0.8 m27, by Zeiss (1 mentions)
Plan apochromat 63x, by Zeiss (1 mentions)
Oct compound, by Sakura Finetek (1 mentions)
Vectashield antifade mounting medium, by Vector Laboratories (1 mentions)
510 meta, by Zeiss (1 mentions)
4 protocols using plan apochromat 10x 0.45 m27
1
Automated CTC Identification by Fluorescence
2
Multimodal Microscopy Systems for Live-Cell Imaging
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Three microscope systems were used for the experiments in this study:
Olympus two-photon microscope system (FVMPE-RS) equipped with a 10× water-immersion objective lens (UMPLFLN10X; numerical aperture (NA), 0.30; working distance (WD), 3.5 mm) and a 25× water-immersion objective lens (XLPLN25XWML; NA, 1.05; WD, 2.0 mm). We used 920 nm and 1040 nm for the excitation of YFP and tdTomato, respectively.
Olympus confocal microscope system (FV1200MPE) equipped with a 10× air objective lens (UPLSAPO10X; NA, 0.4; WD, 3.1 mm), a 20× air objective lens (UCPLFLN20X; NA, 0.7; WD, 0.8 mm), and a 60× oil immersion objective lens (PLAPON60X; NA, 1.4; WD,0.12 mm). We used 405 nm, 488 nm, 594 nm, and 633 nm 1p lasers.
Zeiss confocal microscope system (LSM880) equipped with a 10× air objective lens (Plan-Apochromat 10X/0.45 M27), a 25× glycerol-immersion objective lens (LCI Plan-Neofluar 25x/0.8 Imm Korr DIC M27), a 20× air objective lens (Plan-pochromat 20x/0.8 M27), and a 63× oil immersion objective lens (Plan-Apochromat 63x/1.4 Oil DIC M27). We used 488 nm and 633 nm 1p lasers.
3
Multimodal Imaging of Plant Leaf Anatomy
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Tissue samples of the middle section of adult leaves were collected and fixed in Formalin-Acid-Alcohol (ethanol (>90%) 50 %, glacial acetic acid 5 %, formalin (37% formaldehyde) 10%).
Fixed samples were infiltrated in gradual increases to 30% sucrose solution, embedded in OCT compound (Sakura Finetek USA) and frozen into cryoblocks. 20 µm block face sections were collected on 1% polyethylenimine (PEI) coated slides, stained with 0.1% calcofluor white (aq., Sigma Aldrich) for five minutes followed by 0.01% Fluorol Yellow (Santa Cruz Biotechnology) in lactic acid solution for 30 minutes, mounted in Vectashield anti-fade mounting medium (VECTOR Laboratories), and sealed underneath a coverslip by nail polish. Images were captured on a Zeiss LSM 880 Confocal with FAST Airyscan using Plan-Apochromat 10X/0.45 M27, Plan-Apochromat 20x/0.8 M27, Plan-Apochromat 63x/1.4 Oil DIC M27 objectives set at 515nm emission/488nm excitation wavelength for Fluorol Yellow and 450nm emission/405nm excitation wavelength for calcofluor white. Collected images were processed through superresolution Airyscan and composite pictures were processed through ImageJ. Cell counts of epidermal cell types were done with ImageJ.
Fixed samples were infiltrated in gradual increases to 30% sucrose solution, embedded in OCT compound (Sakura Finetek USA) and frozen into cryoblocks. 20 µm block face sections were collected on 1% polyethylenimine (PEI) coated slides, stained with 0.1% calcofluor white (aq., Sigma Aldrich) for five minutes followed by 0.01% Fluorol Yellow (Santa Cruz Biotechnology) in lactic acid solution for 30 minutes, mounted in Vectashield anti-fade mounting medium (VECTOR Laboratories), and sealed underneath a coverslip by nail polish. Images were captured on a Zeiss LSM 880 Confocal with FAST Airyscan using Plan-Apochromat 10X/0.45 M27, Plan-Apochromat 20x/0.8 M27, Plan-Apochromat 63x/1.4 Oil DIC M27 objectives set at 515nm emission/488nm excitation wavelength for Fluorol Yellow and 450nm emission/405nm excitation wavelength for calcofluor white. Collected images were processed through superresolution Airyscan and composite pictures were processed through ImageJ. Cell counts of epidermal cell types were done with ImageJ.
4
Laser Scanning Microscopy of Nail Imaging
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An inverted laser-scanning microscope (510Meta, Carl Zeiss, Jena, Germany) was used. A HeNe laser (543 nm) was employed to excite NR and the emission signals were collected using a LP 560 filter. Planar (x-y) and cross-sectional confocal images were captured to assess the localization of the fluorophore on or within the nail. Dorsal images were acquired using an oil objective (EC Plan-Neo 40x/1.3 M27, Carl Zeiss). To obtain a direct transverse view of the nail, an air objective (Plan-Apochromat 10x/0.45 M27, Carl Zeiss) was employed. In this case, the treated nail was thinly sectioned and the slices were then glued to a microscope slide with the cross-section orientated towards the objective. All transverse images were recorded from a few microns below the cut surface to avoid any artefact caused by the sectioning. To rule out interference from any nail auto-fluorescence, the laser power and detector settings were minimized so that no signal was detectable from the untreated negative control. Reflectance and/or optical images were captured.
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