All the fluorescence microscopy images of imaginal discs and wing primordia were captured using confocal Leica SP8. Images were recorded at a 1024 × 1024-pixel resolution using oil objective 40×. Expression intensity of GFP, Antp, and Ubx was determined using the histogram function of the FIJI Software. Briefly, threshold was adjusted (using the « Image calculator » function) to create an image containing all positive nuclei (using the « Substract » function) that were analyzed for fluorescence quantification (using the « analyze particles » function) and deduce the mean fluorescence intensity. The adult Drosophila appendage phenotype images were taken by scanning electron microscope Hirox SH-3000 or with a Keyence VHX7000 microscope. The adult wing and haltere were isolated from the rest of the insect body to allow for better manipulation, while mounting samples. Picture of other adult insects were taken with a Keyence VHX7000 microscope.
Vhx 7000 microscope
Manufactured by Keyence
Sourced in Japan
The VHX-7000 is a digital microscope designed for high-resolution imaging and analysis. It features a high-performance CMOS image sensor, advanced optics, and a user-friendly interface. The VHX-7000 enables detailed observation and measurement of a wide range of samples.
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Lab products found in correlation
Live dead viability cytotoxicity kit, by Thermo Fisher Scientific (1 mentions)
Ethidium homodimer 1, by Thermo Fisher Scientific (1 mentions)
Calcein am, by Thermo Fisher Scientific (1 mentions)
Avance nmr spectrometer, by Bruker (1 mentions)
Nicolet is50 ft ir instrument, by Thermo Fisher Scientific (1 mentions)
Dextran sulfate, by Merck Group (1 mentions)
Stereomicroscope, by Keyence (1 mentions)
Nbt bcip, by Roche (1 mentions)
Gotaq polymerase, by Promega (1 mentions)
9 protocols using vhx 7000 microscope
1
Fluorescence Microscopy of Drosophila Appendages
2
Prostate Cancer Stem Cell Culturing and Characterization
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Five to ten million of 22Rv1 and DU145 cells were stained with phycoerythrin (PE)- conjugated anti-CD44 and allophycocyanin-conjugated anti-CD133 antibodies (BioLegend, San Diego, CA, USA), and then, double-stained cells were sorted as the CSC population using an FACS machine (Laguna Hills, Becton-Dickson, CA, USA). A CSC single-cell suspension was split into a low-attachment 6-well plate at 500 cells per well, cultured in serum-free DMEM/F12 media supplemented with 2% B27, 1% N2, 10-ng/ml rhFGF-b, and 20-ng/ml rhEGF, and then treated with 0.01% Dimethyl sulfoxide (DMSO) (vehicle control) or FKA or MLN4924 at indicated concentrations once for 24 h or multiple times by refreshing the treatments every 3 days. After 14 days, spheres were digested with accutase and replated into new plates for culturing for another 14 days without any treatments as the secondary generation; the prostaspheres from the secondary generation were repeated with the same culturing process to evaluate the tertiary generation (10 (link)–12 (link)). The number and diameter of prostaspheres were counted and measured in five independent fields per well under a KEYENCE VHX-7000 microscope at different time points to evaluate the growth of prostaspheres.
3
Microscopic Analysis of Wool Fibre Morphology
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Images of wool fibres, attached to leaves of an isolated D. tapetodes rosette, were taken with a Keyence VHX-7000 microscope at 2500 × magnification and illuminated with full field coaxial light. 2D depth-up mode was used for in-focus acquisitions. Fibre width measurements were carried out using the point-to-point measuring tool in the Keyence software.
4
Bioink Viability Assessment via Live/Dead Assay
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Viability testing was performed using the LIVE/DEAD Viability/Cytotoxicity Kit (Thermo Fisher Inc., Waltham, MA, USA). The bioinks were both pipetted and printed onto a well plate in triplicate. The bioinks were crosslinked and incubated in growth medium for 2 days and 7 days at 37 °C and 5% CO2. After each time point, the growth medium was removed, and the constructs were washed in PBS. A solution containing 2 µM of Calcein AM (Invitrogen, Carlsbad, CA, USA) and 4 µM of ethidium homodimer-1 (Invitrogen) in PBS was added to the wells and incubated for 25 min at 37 °C and 5% CO2. The dye was then removed, and PBS was added to the wells. The constructs were then imaged and overlayed (Keyence VHX-7000 microscope). The images were processed and analyzed using the ImageJ/Fiji software (Version 1.52p, NIH) [30 (link)] and using the StarDist plugin [31 ].
5
Multimodal Characterization of Filaments
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Chemical characterization of the filaments was accomplished with Fourier transform infrared spectroscopy (FTIR), NMR, and laser-induced breakdown spectroscopy (LIBS). Absorption spectra were obtained in attenuated total reflectance (ATR) mode using a Nicolet iS50 FTIR instrument (Thermo Fisher Scientific, Waltham, MA, USA) equipped with a diamond crystal reference. Scans measured the absorbance from 525 to 4000 cm−1 at a resolution of 4 cm−1, and spectra reported here are background-subtracted and averaged from 32 scans per sample. For NMR, samples were dissolved in deuterated chloroform over 48 h, and the resulting solutions were extracted with filtered syringes and placed in NMR tubes. Extracts were analyzed via liquid-state 1H NMR experiments using a Bruker Avance NMR spectrometer (Bruker Corporation, Billerica, MA, USA) operating at 500.13 MHz, and residual protons in the solvent were used as a proton reference. Elemental makeup and distribution of the filaments were determined via a LIBS-based elemental analyzer (Keyence EA-300 VHX Series; Keyence Corporation, Osaka, Japan) attached to a Keyence VHX-7000 microscope. The spot size for LIBS analyses was 10 µm and element concentrations were averaged from three analyses per spot.
6
In Situ Hybridization of Zebrafish Embryos
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In situ hybridization (ISH) probes for MSANTD2 were cloned from WT zebrafish cDNA by PCR using the GoTaq polymerase (Promega). PAX2A and HER5 probes were given by Dr. Sebastian Dworkin lab, La Trobe University, Melbourne, Australia.
Zebrafish embryos were collected, removed from their chorion, sorted and fixed in paraformaldehyde (PFA) 4%, dehydrated in methanol and stored at—20°C. ISH was performed following the Thisse Lab protocol (Thisse and Thisse 2008 (link); 2014 (link)). Embryos were rehydrated and washed in phosphate buffered saline (PBS)—Tween (PBT) solution. They were permeabilized with proteinase K and fixed in PFA. Each embryo was incubated with probes overnight at 65°C in hybridization mix supplemented with 5% Dextran Sulfate (Millipore). Nonhybridized probes were removed with several washes in formamide and saline-sodium-citrate solutions. Embryos were incubated overnight at 4°C with α-DIG (digoxigenin) antibodies (Roche). Nonfixed antibodies were removed with PBT washes. Probes were revealed with nitro blue tetrazolium chloride - 5-bromo-4-chloro-3-indolyl-phosphate (NBT-BCIP) (Roche). Embryos were fixed in PFA. After removing of the background with ethanol bath, embryos were stored in glycerol 80% at 4°C. Pictures were taken under Leica stereomicroscope and Keyence VHX-7000 microscope.
Zebrafish embryos were collected, removed from their chorion, sorted and fixed in paraformaldehyde (PFA) 4%, dehydrated in methanol and stored at—20°C. ISH was performed following the Thisse Lab protocol (Thisse and Thisse 2008 (link); 2014 (link)). Embryos were rehydrated and washed in phosphate buffered saline (PBS)—Tween (PBT) solution. They were permeabilized with proteinase K and fixed in PFA. Each embryo was incubated with probes overnight at 65°C in hybridization mix supplemented with 5% Dextran Sulfate (Millipore). Nonhybridized probes were removed with several washes in formamide and saline-sodium-citrate solutions. Embryos were incubated overnight at 4°C with α-DIG (digoxigenin) antibodies (Roche). Nonfixed antibodies were removed with PBT washes. Probes were revealed with nitro blue tetrazolium chloride - 5-bromo-4-chloro-3-indolyl-phosphate (NBT-BCIP) (Roche). Embryos were fixed in PFA. After removing of the background with ethanol bath, embryos were stored in glycerol 80% at 4°C. Pictures were taken under Leica stereomicroscope and Keyence VHX-7000 microscope.
7
Microscopic Analysis of Mask Layers
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A Keyence VHX-7000 microscope equipped with a 100 × lens was used to observe the microstructure of mask layers. Treated and untreated masks were analyzed to assess the effects of the different treatments on the microstructure of the spunbond and meltblown PP layers.
A scanning electron microscope GeminiSEM from Zeiss was used in High Vacuum mode to observe the structure of a reference and a washed mask. Prior to observation, the samples of individual layers were coated with gold to a thickness of 50 nm.
A scanning electron microscope GeminiSEM from Zeiss was used in High Vacuum mode to observe the structure of a reference and a washed mask. Prior to observation, the samples of individual layers were coated with gold to a thickness of 50 nm.
8
Microscopic Analysis of Sample Structures
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The VHX-7000 microscope (equipped with a 3 MP camera) by Keyence (Osaka, Japan) was used to analyze the structures and fractures of the samples.
9
Histological Analysis of Hearts
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