The grain samples were collected with the same methods as that in 2.2.1 at 3, 5, 7, 9, 11, 13, 15, 18, and 21 DPA. TTC staining and Evans blue staining were used to observe the process and pattern of cell death in endosperm cells. TTC stains viable cells or tissues, while Evans blue dye stains dead cells, which are indicated to be dead by specific stains used in viability assays, such as TTC or Evans blue (van Doorn et al., 2011 (link)). The TTC staining method was modified from Oberle and Watson (1953) . Thin sections of grains were made by hand using sharp double-sided blades at different DPAs (3, 5, 7, 9, 11, 13, 15, 18, and 21 DPA) and stained in 0.5% (w/v) TTC (Aladdin, E104208-10 g, United States) for 30 min at 25°C. Stained sections were photographed with a stereomicroscope (Zeiss Discovery V20, Germany). At least five grains were observed. The Evans blue staining method was modified from Young and Gallie (1999) (link). Grains at 3, 5, 7, 9, 11, 13, 15, 18, and 21 DPA were cut with sharp double-sided blades by hand and stained in 0.1% (w/v) Evans blue (Aladdin, E104208-10 g, United States) for 2 min. Stained sections were washed with water for 1 h and photographed with a stereomicroscope (Zeiss Discovery V20, Germany). At least five grains were observed for each WR and CR.
Discovery v20
Manufactured by Zeiss
Sourced in Germany, United States, Poland
The Discovery V20 is a stereo microscope designed for high-performance observation and documentation. It offers a wide range of magnification options, enabling detailed examination of samples. The microscope features an ergonomic design and intuitive controls for comfortable and efficient use.
Automatically generated - may contain errors
Lab products found in correlation
Inspect s50, by Thermo Fisher Scientific (9 mentions)
Lsm 710, by Zeiss (8 mentions)
Axio imager 2, by Zeiss (8 mentions)
Focus 7, by Helicon (3 mentions)
Axiocam icc5, by Zeiss (3 mentions)
Axiocam mrc5, by Zeiss (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Evo ls15, by Zeiss (2 mentions)
Planapo s fwd 60 mm, by Zeiss (2 mentions)
Axiocam 305, by Zeiss (2 mentions)
Axiocam 705, by Zeiss (2 mentions)
Emitech sc7620, by Quorum Technologies (2 mentions)
Lsm 780, by Zeiss (2 mentions)
Axiocam erc5, by Zeiss (2 mentions)
Lsm 700, by Zeiss (2 mentions)
Axiocam hrc, by Zeiss (2 mentions)
Proteinase k, by Roche (1 mentions)
In situ cell death detection kit, by Roche (1 mentions)
Tmr red, by Roche (1 mentions)
Axiocam 506, by Zeiss (1 mentions)
117 protocols using discovery v20
1
Tracking Endosperm Cell Death in Grains
2
Generating Stable GFP-Kras KH2 ES Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
KH2 ES cells were transfected with LiOnCAG∞GFP-Kras and CAG::hyPBase plasmids (4:1 mass ratio) using Lipofectamine 2000 reagent. 48 hrs after transfection, GFP-positives cells (1.5%) were sorted using an Astrios MoFlo EQ cell sorter and plated at low density (103 cells/ 10-cm dish) on feeder cells. After eight days, GFP-positives clones were picked under a fluorescent stereomicroscope (Zeiss Discovery V20).
3
Fracture Analysis via Stereomicroscopy and SEM
4
Imaging Transgenic Nematodes with Fluorescence
5
DMACA Staining for Plant Phenolic Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The presence of PAs in plant tissue was detected by staining the tissues with DMACA (dimethylaminocinnamaldehyde) solution (1% DMACA, 1% 6 N HCl in methanol). Dried seeds were stained for 6 h and the seedlings for 20 min. After staining, the samples were transferred to distilled water and blue staining was visualized and photographed with a Zeiss Discovery V20 stereomicroscope.
6
Quantifying Dental Adhesion Failure Patterns
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After failure, a blinded examiner, through a stereomicroscope (Discovery V20;
Carl-Zeiss, Gottingen, Germany) at × 40 magnification, evaluated the samples.
Failure patterns were classified into: Ac/d = Predominant adhesive at
cement/dentin interface; Ac/p = Predominant adhesive at cement/post interface; C
= Cohesive of dentin. The calibration consisted of repeating the analysis of the
fracture pattern of 30 slices, with an interval of two weeks. Examiner
reproducibility, which was calculated using the Kappa test, was 0.943.
Carl-Zeiss, Gottingen, Germany) at × 40 magnification, evaluated the samples.
Failure patterns were classified into: Ac/d = Predominant adhesive at
cement/dentin interface; Ac/p = Predominant adhesive at cement/post interface; C
= Cohesive of dentin. The calibration consisted of repeating the analysis of the
fracture pattern of 30 slices, with an interval of two weeks. Examiner
reproducibility, which was calculated using the Kappa test, was 0.943.
7
Embryonic Cell Death Analysis by TUNEL
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Thirty and forty-eight hours post fertilization embryos were fixed with 4% PFA overnight at 4 °C followed by five washes (5 min each) in 1X PBST. Then embryos were treated with acetone for 5 min and digested with proteinase K (Roche) (10 μg/ml) at room temperature for 15–20 min. After each of these treatments, 1X PBST washing was required for next step. TUNEL staining was performed using the in situ Cell Death Detection Kit and TMR Red (Roche). The reaction was stopped after 1.5 h dark treatment and immediately imaged. The pictures were captured by a fluorescence microscope (ZEISS, Discovery.V2.0).
8
Alcian Blue Staining of Shark Embryos
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The fixed embryos subsequently were processed in acid-free alcian blue staining following Walker and Kimmel [45 (link)] with slight modifications. Prior to staining, the bamboo shark embryos were treated with acetone (100%) to degrease the tissue, while a couple of catsharks at stage 31 were stained with alcian blue/acetic acid/ethanol (Ethanol 80%/ Acetic acid 20%) prior to degrease treatment with trypsin. After staining, the bamboo shark embryos were rehydrated through an Ethanol series up to 25%/100 mM Tris pH 7.5, and for the catsharks in a series of Ethanol 25%/PBS. Afterwards, the specimens were bleached in H2O2 3%/KOH 0.5% to remove pigments. Finally, muscles were macerated in 0.5% trypsin/35% sodium borate. The processed embryos were stored in 75% glycerol/0.1% KOH for further studies. All specimens were photographed in ventral view with a Zeiss Discovery V20 stereomicroscope equipped with a Zeiss AxioCam 506 digital camera.
9
Caenorhabditis Nematode Larval Sizing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Twenty-four hours after bleach (~12 hr after hatch), 1000 L1s were pipetted out of the starvation cultures, spun down in 15 ml plastic conical tubes by centrifuge for 1 min at 3000 rpm then plated onto unseeded 10 cm NGM plates. L1s were imaged with a Zeiss Discovery. V20 stereomicroscope at ×77 and measured using Wormsizer (Moore et al., 2013 (link)). Ad libitum concentration was defined as 25 mg/ml and dietary restriction concentration was determined based on what concentration of HB101 produced the largest average L1 size for each strain. For C. elegans, this was 3.13 mg/ml, and eightfold dilution from ad libitum and consistent with previous determinations for dietary restriction in C. elegans (Hibshman et al., 2016 (link)). For C. briggsae, peak L1 size varied between 12.5 and 6.25 mg/ml depending on replicate. We chose to use 6.25 mg/ml as the dietary restriction concentration to be consistent with replicates that were already being processed. The peak L1 size and determination of dietary restriction for C. tropicalis were 6.13 mg/ml. C. kamaaina did not show a significant change in L1 size across conditions and was ultimately excluded from the brood size assay due to difficulty interpreting effects of starvation on brood size in a male–female strain.
10
Ethanol-preserved Mountain Insects of Sichuan
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Adult specimens were captured in the mountainous area of Sichuan province, China in July 2016 and preserved in 75% ethanol. The holotype and paratypes are deposited in the Entomological Museum, Northwest A&F University, Yangling, China (NWAFU
Males and females were dissected and photographs were taken with an advanced Stereo Microscope system Discovery V20 (Zeiss, Germany). Serial photographs were stacked with software Helicon Focus Pro 6.2.2 and further processed with Adobe Photoshop CS6. The measurements of wings were conducted with an electronic digital caliper.
Males and females were dissected and photographs were taken with an advanced Stereo Microscope system Discovery V20 (Zeiss, Germany). Serial photographs were stacked with software Helicon Focus Pro 6.2.2 and further processed with Adobe Photoshop CS6. The measurements of wings were conducted with an electronic digital caliper.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!