Routine analysis of brightfield and fluorescent samples were performed on a Leica DM550B microscope. Peroxidase images were acquired using this microscope equipped with a DFC550 digital camera (Leica). Fluorescent images presented in the figures were acquired using a confocal microscope (Zeiss LSM 700) located at the Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen. Phase contrast images were acquired using a Leica DM IL equipped with a MC170 HD digital camera. For image acquisition at low magnification a Leica MDG41 equipped with a MC170 HD digital camera was used.
Mc170 hd digital camera
Manufactured by Leica
Sourced in Switzerland, Germany
The Leica MC170 HD is a digital camera designed for microscopy applications. It captures high-definition images with a resolution of up to 5-megapixels. The camera features a C-mount interface for easy attachment to microscopes and a built-in CMOS image sensor.
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Lab products found in correlation
Exosome depleted fbs, by System Biosciences (2 mentions)
Labovert fs inverted microscope, by Leica (2 mentions)
Dm1000 microscope, by Leica (2 mentions)
Lsm 700, by Zeiss (2 mentions)
Mz6 stereomicroscope, by Leica (1 mentions)
Dm2000, by Leica (1 mentions)
Dm il led microscope, by Leica (1 mentions)
Application suite las version 4, by Leica (1 mentions)
Dmr microscope, by Leica (1 mentions)
H 7100 transmission electron microscope, by Hitachi (1 mentions)
Automatic microtome, by Leica (1 mentions)
Dm750, by Leica (1 mentions)
Entellan, by Merck Group (1 mentions)
Sz61 stereo microscope, by Leica (1 mentions)
Dfc550 digital camera, by Leica (1 mentions)
Bodipy 493 503, by Thermo Fisher Scientific (1 mentions)
M165 fc, by Leica (1 mentions)
14 protocols using mc170 hd digital camera
1
Integrated Microscopy Imaging Techniques
2
Mouse Tissue β-Galactosidase Staining Protocol
3
Morphometric Analysis of Nematode Specimens
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The extracted nematodes were mounted on temporary microscope slides as reported in Shurtleff and Averre (2000) and morpho-biometrically characterized. Specimens were observed under a light microscope Leica DM2000 equipped with MC 170 HD digital camera using LEICA Application Suite v. 4.9.0 as measuring software (Leica Microsystems, Heerbrugg, Switzerland). Morphological characters were selected according to Goodey (1963) and Meamar et al. (2007) (link). The measures were taken from adult specimens (14 females and 21 males) collected from P. archon larvae of the second palm. The features were body length (L), maximum body diameter (MBD), buccal cavity length (B), tail length (T), and distance of the base of the pharynx from the head end (ES). In addition, distance of vulva from head end (V) and egg diameter (ED) were measured in females, while spicule length (SP) and gubernaculum length (GU) were measured in males. The following ratios were then calculated: a (L/MBD), b (L/ES), c (L/T), and V% (V/L*100).
4
Isolation and Culture of Human Cardiac Progenitor Cells
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Human cardiac progenitor cells (hCPCs) were isolated from the right atrial appendage and sorted for expression of c-kit cell surface marker, as described previously (Zhang et al., 2017 (link)). Cells were used at passage 7–10 for these studies. Cells were initially cultured for 48 h at normoxic conditions (37°C, 21% O2) then placed in medium with exosome-depleted FBS (SBI, Palo Alto, CA, United States) and continuously cultured at normoxic condition of 21% O2 physoxic condition of 5% O2 or hypoxic condition of 1% O2 using a controlled C-chamber incubator (ProOx P110 O2 Controllers, BioSperix, Parish, NY, United States). Media was refreshed every other day, retaining 20% of conditioned media. Phase-contrast images were captured using a DM IL LED microscope and MC170 HD digital camera (Leica Microsystems, Inc., Buffalo Grove, IL, United States).
5
Macroscopic Analyses of Plant Material
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Macroscopic analyses were executed according to the methods of the European Pharmacopoeia (10th edition, 2019) [64 ]. The plant material was analyzed by naked eye and with an Olympus SZ61 stereomicroscope (Leica Microsystems Ltd., Heerbrugg, Switzerland) coupled with a Leica MC170 HD digital camera controlled by Leica Application Suite (LAS) Version 4.8.0 software (Leica Microsystems Ltd., Heerbrugg, Switzerland).
6
Scratch Assay on hAECs with Refeed® Supplement
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Scratch assay was performed on hAECs from P1 to P4 in both basal medium and Refeed® supplemented one. Cells, seeded at 60,000 cells/cm2 in a 24-well culture plate, were allowed to grow for 24 h to obtain a confluent monolayer. The scratch was made using plastic sterile tips, and then, wells were washed with PBS to remove detached cells. Fresh culture medium with or without Refeed® was added to the wells. Scratched monolayers were monitored, and images were acquired using an optical microscope Leica Labovert FS inverted Microscope (Leica Microsystems, Wetzlar, Germany) with a Leica MC170 HD digital camera (Leica Microsystems, Wetzlar, Germany) at regular interval times until their complete closure. Scratch areas were measured with the NIH ImageJ program [38 (link)] (pp. 671–675), and the percentage of scratch closure was calculated using GraphPad Prism software. The results are represented as the percentage of uncovered area ± SD. The scratch area at T0 was set as 100% for both culture conditions.
7
Quantifying Cellular Senescence via SA-β-Gal Assay
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The Senescence-Associated β-Galactosidase assay (SA-β-Gal) was performed on hAECs cultured from P1 to P4. Cells were seeded in 24-well culture plates (5000 cells/cm2) and cultured in basal medium with or without Refeed®. The SA-β-Gal assay was performed using a commercial kit (Cell Signaling Technology, Danvers, MA, USA) following the manufactured instructions. The staining reaction was stopped after six hours; images of stained hAECs were acquired using Leica Labovert FS inverted Microscope (Leica Microsystems, Wetzlar, Germany) with Leica MC170 HD digital camera (Leica Microsystems, Wetzlar, Germany), and plates were stored at 4 °C in 70% glycerol. The number of blue-stained SA-β-Gal-positive cells in 10 random fields of the wells was counted; data were analyzed using GraphPad Prism 7.04 software. Results are represented as the percentage of blue-stained cells (SA-β-Gal positive cells) ± SD at each passage for both conditions.
8
Bursaphelenchus Specimen Preparation and Measurement
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Bursaphelenchus specimens were collected from the Petri dish, washed in sterilized water and heat-killed in warm water at 65°C; then they were fixed in triethanol-amine-formalin (Courtney et al., 1955 ), processed in glycerin by a modification of the glycerin-ethanol series of Seinhorst (1959) rapid method and finally, permanently mounted in anhydrous glycerin on glass microscope slides.
In total, 10 females and 10 males were photographed and measured. Photographs were taken with a Leica DM2000 light microscope using a Leica MC170 HD digital camera (Leica, Heerbrugg, Switzerland). Measurements were performed with the LEICA Application Suite (LAS) Version 4.9.0. Morphological and molecular characteristics were compared with the original description (Franklin and Hooper, 1962 ).
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Bursaphelenchus specimens were collected from the Petri dish, washed in sterilized water and heat-killed in warm water at 65°C; then they were fixed in triethanol-amine-formalin (Courtney et al., 1955 ), processed in glycerin by a modification of the glycerin-ethanol series of Seinhorst (1959) rapid method and finally, permanently mounted in anhydrous glycerin on glass microscope slides.
In total, 10 females and 10 males were photographed and measured. Photographs were taken with a Leica DM2000 light microscope using a Leica MC170 HD digital camera (Leica, Heerbrugg, Switzerland). Measurements were performed with the LEICA Application Suite (LAS) Version 4.9.0. Morphological and molecular characteristics were compared with the original description (Franklin and Hooper, 1962 ).
9
Transmission Electron Microscopy of Bacterial Cultures
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Pure cultures were processed for transmission electron microscopy as described previously 73 with slight modifications. Cultures were fixed in 3% glutaraldehyde, 1% fresh formaldehyde, and 0.75% tannic acid in 0.05 M Na-cacodylate buffer, pH 7, for 3 h at room temperature. After several rinses in 0.1M buffer, the samples were post-fixed in buffered (0.1M, pH 6.8) 1% osmium tetroxide overnight at 4 C, dehydrated in an ethanol series and embedded in Spurr's resin via ethanol. Thin sections were cut with a diamond histo-knife, stained with methanolic uranyl acetate for 15 min and in Reynolds' lead citrate for 10 min, and observed with a Hitachi H-7100 transmission electron microscope at the Imaging-Microscopy Platform of the IBIS, Universite Laval. Images of colonies on an agar plate were taken by a SMZ-171TLED stereomicroscope (Bio Pioneer Tech Co, Taiwan) equipped with a TrueChrome AF digital camera (TUCSEN, China). Light microscopy images were taken under a Leica DMR microscope (Leica, Germany) equipped with a Leica MC 170HD digital camera (Leica, Germany).
10
Histological Analysis of Muscle Tissue
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The muscle samples (biceps) were removed and fixed in formaldehyde. After fixation, the tissues were dehydrated in ethanol and embedded in resin containing Historesin® hardener (Leica, Wetzlar, HE, Germany). Histological sections were obtained using an automatic microtome (Leica). They were then submitted to staining using the hematoxylin and eosin (H&E) technique, mounted with Entellan (Merck), and analyzed under a light microscope (Leica DM750).
The images of the histological sections were obtained with a 20× objective using the LEICA MC170 HD digital camera. From 10 photos per histology slide, the areas containing muscle fiber, connective tissue (which was considered fibrosis when the quantity of collagen was exacerbated), and inflammatory infiltrate in the muscle were quantified using the ImagePro-Plus® application version 4.5 (Media Cybernetics, Rockville, MD, USA) by manually counting points on the tissue [39 (link)].
The images of the histological sections were obtained with a 20× objective using the LEICA MC170 HD digital camera. From 10 photos per histology slide, the areas containing muscle fiber, connective tissue (which was considered fibrosis when the quantity of collagen was exacerbated), and inflammatory infiltrate in the muscle were quantified using the ImagePro-Plus® application version 4.5 (Media Cybernetics, Rockville, MD, USA) by manually counting points on the tissue [39 (link)].
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