The size and morphology of the PLGA implants were studied using an Olympus SZX9 microscope with a UC30 camera (Olympus Optical Co., Hamburg, Germany) and OLYMPUS stream motion software (Olympus Optical Co., Hamburg, Germany). The length of each implant was adjusted to 3.0 mm or 5.0 mm by cutting the extrudates under the microscope with a scalpel. A tolerance of 0.1 mm was accepted.
Uc30 camera
Manufactured by Olympus
Sourced in Japan, Germany
The UC30 camera is a compact, high-resolution microscope camera designed for laboratory applications. It features a 3.1-megapixel CMOS sensor and can capture images and videos at various resolutions. The camera is compatible with a wide range of microscopes and can be easily integrated into existing imaging systems.
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Lab products found in correlation
Bx51 fluorescence microscope, by Olympus (3 mentions)
Cx 41 1, by Olympus (3 mentions)
Sz61 stereomicroscope, by Olympus (3 mentions)
Victor x3, by PerkinElmer (3 mentions)
Xc30 microscope, by Olympus (2 mentions)
Image pro software, by Media Cybernetics (2 mentions)
300 cp system, by EXAKT (1 mentions)
Szx7 stereomicroscope, by Olympus (1 mentions)
Bx45 microscope, by Olympus (1 mentions)
Analysis pro software, by Olympus (1 mentions)
Ckx41 inverted microscope, by Olympus (1 mentions)
Bx43f light microscope, by Olympus (1 mentions)
U cmad3 adapter, by Olympus (1 mentions)
Laminin, by Merck Group (1 mentions)
Neurobasal plus medium, by Thermo Fisher Scientific (1 mentions)
B 27 plus supplement, by Thermo Fisher Scientific (1 mentions)
Cultureone supplement, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Poly l ornithine, by Merck Group (1 mentions)
30 protocols using uc30 camera
1
PLGA Implant Size and Morphology
2
PLGA Implant Swelling Characterization
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For the visualization of the prepared extrudates two different microscopes were used. Size and swelling behavior of PLGA implants were studied using an Olympus SZX9 reflected-light microscope.
Samples (n = 3) were prepared by cutting the extrudates under the microscope to a length of 3.0 mm. The swelling of the extrudates was monitored using 1 mL glass vials filled with 1 mL phosphate buffer pH 7.4. The vials were kept at 37 °C in a shaker with light protection (Memmert GmbH + Co. KG, Schwabach, Germany). The extrudates were carefully removed at pre-determined time points and examined microscopically. Pictures were taken with an UC30 camera (Olympus Optical Co., Hamburg, Germany) and the dimensions of the implants were analyzed with OLYMPUS stream motion (Olympus Optical Co., Hamburg, Germany).
The presence of dexamethasone crystals was examined with an Axiolab transmitted-light microscope with polarization filter unit (Carl Zeiss MicroImaging GmbH, Göttingen, Germany). For this purpose, the extrudates were cut in 30 µm slices using a Leica RM 2245 microtome (Leica Biosystems, Nussloch, Germany).
Samples (n = 3) were prepared by cutting the extrudates under the microscope to a length of 3.0 mm. The swelling of the extrudates was monitored using 1 mL glass vials filled with 1 mL phosphate buffer pH 7.4. The vials were kept at 37 °C in a shaker with light protection (Memmert GmbH + Co. KG, Schwabach, Germany). The extrudates were carefully removed at pre-determined time points and examined microscopically. Pictures were taken with an UC30 camera (Olympus Optical Co., Hamburg, Germany) and the dimensions of the implants were analyzed with OLYMPUS stream motion (Olympus Optical Co., Hamburg, Germany).
The presence of dexamethasone crystals was examined with an Axiolab transmitted-light microscope with polarization filter unit (Carl Zeiss MicroImaging GmbH, Göttingen, Germany). For this purpose, the extrudates were cut in 30 µm slices using a Leica RM 2245 microtome (Leica Biosystems, Nussloch, Germany).
3
Visualizing Apoptosis via Nuclear Condensation
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The nuclear condensation was examined using the cell-permeable DNA dye Hoechst 33342. For this, HeLa and SiHa cells were plated (2.5 × 105 cells/mL) in a 24-well culture plate (500 μL/well) containing a sterile round coverslip and incubated at 37 °C with 5% CO2 for 24 h. The cells were then treated with A3K2A3 (IC50 and 2xIC50 according to cell line) for 48 h. The PC was carried out with CPT (10 μM). The cells were washed with PBS, labeled with 10 μg/mL Hoechst 33342, incubated for 20 min in the dark at 37 °C, and then washed again twice with PBS. They were then visualized on a fluorescence microscope (Olympus®, BX51) and the images were captured with an Olympus® UC30 camera. The identification criteria were living cells with homogeneously stained nuclei (light blue), while cells strongly stained with an intense and bright blue color were indicative of apoptosis due to chromatin condensation. The images were analyzed using ImageJ software [32 (link)].
4
Histopathological Analysis of Acupuncture Needle Sites
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Acupuncture needles were removed and the surrounding tissue was explanted and fixed in 10% neutral-buffered formalin (100 ml 40% formalin, 900 ml ddH2O, 4 g l−1 NaH2PO4 and 6.5 g l−1 Na2HPO4, pH 7). The tissue samples were trimmed, dehydrated in increasing concentrations of ethanol, cleared with xylene, infiltrated and embedded in paraffin wax, sectioned at 2–4-µm thickness using an EXAKT 300 CP system (EXAKT Technologies) and stained with hematoxylin and eosin. The tissue sections were analyzed by light microscopy and images were acquired with an Olympus UC30 camera. The tissue around the acupuncture needle footprint was histopathologically evaluated by a pathologist at AnaPath Services according to the ISO 10993-6:2016(E) standard. In addition, collagen denaturation was scored to exclude any potential thermoelectrical impact on the tissues following electrostimulation.
5
Analysis of DNA Damage in Epimastigotes
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DNA double-strand ruptures were analyzed in situ using a TUNEL kit. Epimastigotes (1 × 106 cells/mL) were treated with 18.0 and 77.0 μM of C4 for 24 h at 28°C, after the cells were subjected to the TUNEL assay according to the manufacturer’s instructions. Actinomycin D (10.0 μg/mL) was used as a positive control. Fluorescence was observed in an Olympus BX51 fluorescence microscope, and pictures were captured with an Olympus UC30 camera.
6
Microscopic Analysis of Fungal Antagonism
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Dual cultures of F. culmorum KF846 with Trichoderma or Clonostachys strains were used for microscopic observations. They were grown on a sterile strip of 20 mm cellophane membrane (50-μm thick) placed before fungal inoculation on a PDA medium in the middle of 8.5 cm Petri dish. Fungi were inoculated on opposite sides of Petri dishes at a distance of 5 mm from the edge. After 7 and 14 days of incubation at 25 ± 2 °C through 12 h/12 h of darkness/light, the mycelium overgrown cellophane membrane was cut with a razor blade in sterile conditions, placed on a microscope slide in a drop of distilled water and examined. Observations were carried out using a light microscope (Olympus CX-41-1 with UC-30 camera, Olympus Corporation, Tokyo, Japan). The samples were screened for loops of the Trichoderma or Clonostachys around F. culmorum hyphae and the anatomical damage of the pathogen.
7
Investigating the Impact of Snail Mucus on Tardigrade Tuns
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After 7 days of anhydrobiosis, one individual of C. nemoralis was transferred to each dish with tardigrade tuns and was left there for 1 min allowing the snail to actively crawl over the tuns. 30 min after the snail was removed from the dish, tardigrade tuns were observed under the Olympus SZX7 stereomicroscope for any animal movements. Then, all covered and vented dishes were left in the Q-Cell incubator overnight. After 24 h, the dried tuns were rehydrated by adding 3 ml of water to each Petri dish to check whether snail’s mucus affected mortality rates of tardigrades. After 3 and 24 h following rehydration tardigrade tuns were observed for any animal movements. Pictures of tuns were taken using Olympus SZ61 stereomicroscope associated with Olympus UC30 camera (Fig. 3 ). As reference data on the rehydration of the M. inceptum tuns free of the snail’s mucus, we used the data from Roszkowska et al.20 who tested anhydrobiosis survivability of above-mentioned species. Individuals used for the tuns preparation in the control option were collected from the same laboratory breeding stock, and prepared at the same laboratory conditions as those used in our experiments20 .![]()
Milnesium inceptum tuns: (
8
Rat Liver Histopathological Analysis
9
Histopathological Cartilage Thickness Measurement
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Immediately after MRI, the carpal joints were opened by sharp dissection carefully to avoid any damage to the cartilage. Osteochondral wedge sections (5 × 5 × 10 mm) were cut out using an oscillating saw. The samples were fixed in formaldehyde, decalcified in formic acid, embedded in paraffin, cut into 2 μm sections and stained with hematoxylin‐eosin. All samples were sliced in the sagittal plane. Images of the sections were obtained using an Olympus BX45 microscope and an Olympus UC30 camera and processed with Olympus cellSens Entry 2.1 software (Figure 2 ). The cartilage thickness was measured with the “Arbitrary Distance” tool with an accuracy of 0.01 μm (Figure 6 ). The histopathology observer was blinded to the US and MRI measurements. Histopathology measurements were performed by one of the authors (CA) after thorough training and alignment by and with a Diplomate of the European College of Veterinary Pathologists. The thickness of the noncalcified cartilage (NCC) and the total cartilage thickness (the NCC plus the calcified cartilage zone (CCZ)) were each measured three times for every location (Figure 6 ). The mean ± SEM of the three repetitions was used for comparison of cartilage thickness with the other modalities.
10
Histopathological Evaluation of Biomedical Samples
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Histopathology evaluation was performed by AnaPath GmbH using a scoring system according to ISO 10993-6:2016(E). The images were taken by an Olympus UC30 camera.
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