RNA in situ hybridization was performed on 3 µm thick formalin-fixed paraffin-embedded sections of rat kidneys using the RNAscope 2.5 HD assay-brown (Advanced Cell Diagnostics) according to the user manuals 322452-USM and 322310-USM using standard conditions. EPO-mRNA was detected by the RNAScope probe Rn-Epo (Cat No. 455901) followed by semi-quantitative scoring, calculating the average count of EPO producing cells per field of view in the kidney cortex and medulla. In situ hybridization was performed with n = 8 kidneys in each group. For determination of EPO-positive cells 10 randomly chosen FOV per renal cortex or 5 randomly chosen FOV per medulla were microscopically analysed and counted [Aperio CS2 (Leica Biosystems), objective: UPlanSApo 20x/0.75 (Olympus)].
Uplansapo 20x 0
Manufactured by Olympus
Sourced in Japan
The UPlanSApo 20x/0.75 is a high-numerical aperture objective lens designed for use in various microscopy applications. It provides a magnification of 20x and a numerical aperture of 0.75, allowing for high-resolution imaging and excellent light-gathering capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Aperio cs2, by Leica (1 mentions)
Rnascope 2.5 hd assay brown, by Advanced Cell Diagnostics (1 mentions)
Ix83 inverted microscope, by Olympus (1 mentions)
Doc cam hr ccd camera, by Molecular Devices (1 mentions)
Ix3 zdc laser based autofocusing system, by Olympus (1 mentions)
Spectra x light engine, by IDEX Corporation (1 mentions)
Ff458 di02 25x36, by IDEX Corporation (1 mentions)
Ff01 483 32 25, by IDEX Corporation (1 mentions)
Ff01 542 27 25, by IDEX Corporation (1 mentions)
Stage top incubator, by Tokai Hit (1 mentions)
3 protocols using uplansapo 20x 0
1
Quantification of Renal EPO Expression
2
Localized Heat Shock in Aspergillus nidulans
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Aspergillus nidulans cells were grown on minimal medium agar plates at 30°C for 2 days. Then, the edges of colonies were cut into 5 mm squares and placed in glass dishes for observation under an inverted microscope IX-83 (Olympus). A localized heat shock was imposed on individual hypha using an IR-LEGO 1000 system (Sigma Koki) (54 (link), 55 (link)), equipped with custom-made UPlanSApo 20x/0.75 and UPlanSApo 40x/0.95 objective lens (Olympus), operating at 15 mW for 5–20 s. MetaMorph and Image J software were used for image analysis.
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Aspergillus nidulans cells were grown on minimal medium agar plates at 30°C for 2 days. Then, the edges of colonies were cut into 5 mm squares and placed in glass dishes for observation under an inverted microscope IX-83 (Olympus). A localized heat shock was imposed on individual hypha using an IR-LEGO 1000 system (Sigma Koki) (54 (link), 55 (link)), equipped with custom-made UPlanSApo 20x/0.75 and UPlanSApo 40x/0.95 objective lens (Olympus), operating at 15 mW for 5–20 s. MetaMorph and Image J software were used for image analysis.
3
FRET Imaging Protocols for Live Cell Analysis
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FRET images were obtained and processed under essentially the same conditions and procedures as previously described (Aoki and Matsuda, 2009) . Briefly, cells were imaged with an IX83 inverted microscope (Olympus, Tokyo, Japan) equipped with a UPlanFL-PH 10x/0.3 (Olympus), a UPlanSApo 20x/0.75 (Olympus), or a UPlanSApo 40x/0.95 objective lens (Olympus), a DOC CAM-HR CCD camera (Molecular Devices, Sunnyvale, CA), a Spectra-X light engine (Lumencor Inc., Beaverton, OR), an IX3-ZDC laser-based autofocusing system (Olympus), an electric XY stage (Sigma Koki, Tokyo, Japan), and a stage top incubator (Tokai Hit, Fujinomiya, Japan). The filters and dichromatic mirrors used for time-lapse imaging were as follows: for FRET imaging, a 438/24 excitation filter (incorporated in the Spectra-X light engine), a FF458-Di02-25x36 (Semrock, Rochester, NY) dichromatic mirror, and FF01-483/32-25 (Semrock) and FF01-542/27-25 (Semrock) emission filter for CFP and FRET, respectively. For mCherry and mScarlet-I imaging, a 575/25 excitation filter, a glass dichromatic mirror (Olympus), and FF01-624/40-25 (Semrock) emission filters were used.
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