For histopathology, skin tissue was fixed in 10% formalin, followed by 70% ethanol and embedded in paraffin, sectioned, and stained with H&E by UCSF Mouse Pathology Core. The inflammatory response following tape-stripping was scored as follows: three parameters (overlying crust, dermal neutrophils, and neutrophilic fat infiltration) were scored separately and then added together. For scoring of crust formation, 0, no crust; 1, little crust present; 2, severe crust formation; for scoring of dermal neutrophils, 0, none present; 1, scarce infiltrate; 2, moderate infiltrate; 3, abundant infiltrate; for scoring of subcutaneous fat infiltration, 0, healthy fat; 1, scarce neutrophilic infiltrate; 2, moderate infiltrate; 3, dense infiltrate partially obliterating fat architecture. Scores were independently corroborated by two reviewers who were unblinded to the experimental set-up. Slides were digitally imaged at 20x with the Aperio AT2 scanner (Leica Biosystems, Vista, CA) using a 20x/0.75NA Plan Apo objective with a 2x optical mag changer.
Plan apo objective
Manufactured by Leica
Sourced in United States
The Plan Apo objective is a high-performance microscope objective lens designed by Leica. It provides excellent image quality and resolution, with minimal distortion and aberrations across the entire field of view. The lens is optimized for applications requiring precise and accurate imaging.
Automatically generated - may contain errors
Lab products found in correlation
Aperio at2, by Leica (2 mentions)
Lex2 b, by BrainVision (2 mentions)
Ff01 543 50 25, by IDEX Corporation (2 mentions)
1.0 plan apo objective, by Leica (2 mentions)
Aperio at2 scanner, by Leica (1 mentions)
Wavefx, by Leica (1 mentions)
Orca r2, by Hamamatsu Photonics (1 mentions)
Dmi6000 microscope, by Leica (1 mentions)
Csu x1 spinning disk, by Yokogawa (1 mentions)
Plan apochromat objective, by Zeiss (1 mentions)
Orca flash4.0 lt cmos camera, by Hamamatsu Photonics (1 mentions)
Lsm 880 with airyscan, by Zeiss (1 mentions)
Zyla 4.2 plus scmos camera, by Oxford Instruments (1 mentions)
Axio observer z1 microscope, by Zeiss (1 mentions)
Axioplan2 wide field microscope, by Zeiss (1 mentions)
Stedycon, by Abberior (1 mentions)
7 protocols using plan apo objective
1
Histopathological Evaluation of Inflammatory Response
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Whole Slide Scanning and Viewing Workflow
3
Whole Slide Scanning and Viewing Workflow
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Glass slides were scanned using Leica Aperio AT2 (Leica Biosystems, Buffalo Grove, Illinois, USA) whole slide scanners at ×40 (0.25 μm/pixel) equivalent magnification using a 20×/0.75NA Plan Apo objective and ×2 automatic optical magnification changer. WSIs are accessed through the PICSPlus module in CoPathPlus and launched into the MSK Slide Viewer, an internally developed whole slide scanner, vendor agnostic whole slide image viewer.12 (link) The MSK Slide Viewer allows for standard whole image viewing, slide label viewing, zooming, panning, and slide navigation. The viewer has tools for manual annotation including a ruler for measurements, tracking of viewed slide regions, screenshots, and comments.12 (link)
4
Widefield Imaging and Retinotopic Mapping
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For widefield imaging we used a standard epi-illumination imaging system35 (link),36 (link) together with an SCMOS camera (pco.edge, PCO AG). A Leica 1.6x Plan APO objective was placed above the imaging window and a custom black cone surrounding the objective was fixed on top of the headplate to prevent contamination from the monitors’ light. The excitation light beam emitted by a high-power LED (465 nm LEX2-B, Brain Vision) was directed onto the imaging window by a dichroic mirror designed to reflect blue light. Emitted fluorescence passed through the same dichroic mirror and was then selectively transmitted by an emission filter (FF01-543/50-25, Semrock) before being focused by another objective (Leica 1.0 Plan APO objective) and finally detected by the camera. Images of 200 x 180 pixels, corresponding to an area of 6.0 x 5.4 mm were acquired at 50 Hz.
To measure retinotopy we presented a 14°-wide vertical window containing a vertical grating (spatial frequency 0.15 cycles/°), and swept37 (link),38 (link) the horizontal position of the window over 135° of azimuth angle, at a frequency of 2 Hz. Stimuli lasted 4 s and were repeated 20 times (10 in each direction). We obtained maps for preferred azimuth by combining responses to the 2 stimuli moving in opposite direction, as previously described37 (link).
To measure retinotopy we presented a 14°-wide vertical window containing a vertical grating (spatial frequency 0.15 cycles/°), and swept37 (link),38 (link) the horizontal position of the window over 135° of azimuth angle, at a frequency of 2 Hz. Stimuli lasted 4 s and were repeated 20 times (10 in each direction). We obtained maps for preferred azimuth by combining responses to the 2 stimuli moving in opposite direction, as previously described37 (link).
5
Widefield Imaging and Retinotopic Mapping
6
Spinning Disk Confocal Microscopy Imaging
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Slides were observed using a Quorum Wave-FX spinning disk confocal microscope and a Leica 63X oil immersion Plan-apo objective (NA 1.4). The solid state lasers used were 491 nm and 561 nm. The emission filters used were bandpass 525/50 nm and bandpass 620/60 nm respectively. The CCD camera used to capture images was a Hamamatsu Orca R2. The software used for image capture and analysis were Metamorph version 7.7.9.0 and Velocity version 6.1.1. Photoshop CS2 was used to label images and create figure montages.
7
High-Resolution Imaging Across Scales
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Widefield imaging was performed using an Axioplan 2 widefield microscope (Zeiss) with 20x NA 0.8 Plan-Apochromat and 100x NA 1.4 Plan-Neofluar objectives, ET filter cubes (Chroma), LED illuminator (Lumencor SOLA), and Coolsnap HQ (Photometrics).
Confocal images were acquired using a CSU-X1 spinning disk (Yokogawa) system mounted on a DMI6000 microscope (Leica) with 100× NA 1.4 HC PLAN APO CS2 objective, 4-channel solid-state Spectral-Borealis laser launch (Andor), a Zyla 4.2 Plus sCMOS camera (Andor). A STEDYCON (Abberior) system mounted on a Zeiss Axio observer Z1 microscope equipped with a 100X NA 1.46 α Plan-Apochromat objective (Zeiss) was used in laserscanning confocal mode, and a Zeiss LSM 880 with Airyscan equipped with a 63X/NA 1.40 Plan-Apochromat objective (Zeiss) was used in super resolution mode with 0.16 μm/pixel, 1024 × 1024 frame size, 2× averaging, and 4x digital zoom.
Tiled tissue microarray images (widefield) were collected using a DM4000 widefield microscope with 20X NA 0.7 HC PLAN APO objective (Leica), ORCA-Flash 4.0 LT+ CMOS camera (Hamamatsu), motorized emission filter wheel and xyz stage (Ludl), 5-channel Aura light engine (Lumencor), and multichannel dichroic matched to single band excitation filters (Semrock), as described (49 (link)).
Confocal images were acquired using a CSU-X1 spinning disk (Yokogawa) system mounted on a DMI6000 microscope (Leica) with 100× NA 1.4 HC PLAN APO CS2 objective, 4-channel solid-state Spectral-Borealis laser launch (Andor), a Zyla 4.2 Plus sCMOS camera (Andor). A STEDYCON (Abberior) system mounted on a Zeiss Axio observer Z1 microscope equipped with a 100X NA 1.46 α Plan-Apochromat objective (Zeiss) was used in laserscanning confocal mode, and a Zeiss LSM 880 with Airyscan equipped with a 63X/NA 1.40 Plan-Apochromat objective (Zeiss) was used in super resolution mode with 0.16 μm/pixel, 1024 × 1024 frame size, 2× averaging, and 4x digital zoom.
Tiled tissue microarray images (widefield) were collected using a DM4000 widefield microscope with 20X NA 0.7 HC PLAN APO objective (Leica), ORCA-Flash 4.0 LT+ CMOS camera (Hamamatsu), motorized emission filter wheel and xyz stage (Ludl), 5-channel Aura light engine (Lumencor), and multichannel dichroic matched to single band excitation filters (Semrock), as described (49 (link)).
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