The histological observation was performed by staining with hematoxylin/eosin (H&E), Giemsa, and trichrome stains using paraffin sections. For H&E staining, paraffin-embedded sample slides were de-paraffinized, hydrated, and then stained with hematoxylin for 1 min. After rinse, the slides were stained with eosin for 5 min, rinsed, and sealed with cover slips. Tissue sections from tumor mass, kidneys, spleen, liver, heart, and lungs were used. The slides were counterstained with hematoxylin and mounted. To determine the effect of CPT on expression of Ki67, PCNA, Caspase-3, Caspase-8, Caspase-9, Bcl-2, Bax, Bid, Bad, Drp1, Opa1, Mfn1, and Mfn2 by immunohistochemistry, the slides were blocked in 5% bovine serum for 15 min, followed by incubation with the primary antibody at 4 °C overnight in a moist chamber. The sections were then incubated with the corresponding secondary antibodies. The antigen-antibody complex was detected by Dako Liquid DAB + Substrate-Chromogen System (Dako, Carpinteria, CA). All slides were examined under light microscopy. Giemsa stain and trichrome stain were performed according to the manufacturer’s instruction of Giemsa Stain Kit (ab150670) and Trichrome Stain Kit (ab150686) from Abcam (UK). All slides were examined under light microscopy.
Giemsa stain kit
Manufactured by Abcam
Sourced in United Kingdom, China, United States
The Giemsa Stain Kit is a laboratory product that provides a solution for staining cells and tissues. It is a widely used staining method in various applications, including hematology, parasitology, and cytology. The kit contains the necessary components to perform the Giemsa staining procedure, which is a versatile and reliable technique for differentiating and visualizing cellular structures and components.
Automatically generated - may contain errors
8 protocols using giemsa stain kit
1
Histological and Immunohistochemical Evaluation of Tumor Samples
2
Colony Formation Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell with indicated treatment were trypsinized and resuspended in culture medium. Cells were seeded into a 6-well plate (1000 cells/well) and cultured for 14 days, and the culture medium was changed every 3 days during the period. After 14 days, cells were fixed with 4% paraformaldehyde at room temperature for 10 mins and stained with Giemsa reagent (Giemsa Stain Kit, Abcam ab150670) or 0.5% crystal violet (Beyotime, Shanghai, China) for 20 mins. Subsequently, the number of colonies was counted and the morphology of the colonies was photographed under Leica AM6000 microscope (Leica, Wetzlar, Germany).
3
Giemsa Staining of TAMs and TAMEMs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To observe the cell morphology of TAMs, TAMEMs, and other cells, Giemsa staining was performed according to the manufacturers' protocols (Abcam, Giemsa Stain Kit). Briefly, spread cells on glass slides and slides were then air‐dried for 10 min prior to staining. Immerse slides in Giemsa staining solution for 10–15 min. After removing the excess staining, soak the slides in PBS buffer until no excess staining solution flows out. Before taking pictures with the microscope, wipe the back of the slide in a vertical position and set it to dry and xylene clear for 10 min.
4
Quantitative Colony Formation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PC-3 cells or PC-3-si-LINC01207 cells were seeded into a 6-well plate at a density of 500 cells/well after cell counting. The culture medium was changed every 2 days. After 14 days of culture, the cells were fixed in 100% methanol for 30 min at room temperature and stained for 10 min using Giemsa Stain kit (Abcam; cat. no. ab150670) according the manufacturer's instructions. After 14 days, an image of the cell colony was captured with a digital camera using an AM6000 microscope (Leica Microsystems, Inc.). Colonies consisting of >50 cells were counted manually.
5
Evaluating Long-Term Proliferation Potential
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Colony formation assay was used to evaluate the long-term proliferation potential. Cells were infected with recombinant lentivirus carrying shRNA or co-transfected with corresponding microRNA mimics, inhibitor or siRNA, as mentioned in the shRNA and siRNA expression Sect. 48 h post-transfection, cells were seeded into a 6-well plate at the density of 200 cells/well after cell counting. The culture medium was changed every two days. After two weeks’ culture, cells were fixed with paraformaldehyde and stained using Giemsa Stain Kit (Abcam, ab150670) according to the manufacturer’s instructions. Finally, the number of colonies formed in each condition was counted using Leica AM6000 microscope.
6
Colony Formation Assay for PC-3 and LNCaP Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PC-3 and LNCaP cells with different treatment were resuspended in fresh medium and then seeded into the 6-well-plates (1000 cells/well). The cells were grown under the condition of 37 °C and 5% CO2 for 14 days, and the medium was replaced every three days. Cells were fixed by 4% paraformaldehyde for 20 min, and stained using Giemsa Stain Kit (Abcam, ab150670) for 20 min at room temperature. Colony morphology were photographed and colony number was counted under Leica AM6000 microscope [41 (link)]. The colony formation assay was conducted with three independent biological replicates.
7
Colony Formation Assay for Cell Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Colony formation assay was used to evaluate the long-term proliferation potential of HT-29 and HCT116 cells. Cells were infected with recombinant lentivirus carrying shRNA targeting lncRNA DCST1-AS1 or co-transfected with miR-582-5p-inhibitor (inh) or pcDNA3.1-HMGB1 or their corresponding controls. Forty-eight hours post-transfection, cells were seeded into a 6-well plate at the density of 200 cells/well. The culture medium was changed every two days. After 14 days’ culture, cells were fixed with 4% paraformaldehyde and stained using Giemsa Stain Kit (Abcam, ab150670) according the manufacturer’s instructions. Finally, the number of colonies formed in each condition was counted using Leica AM6000 microscope [36 (link)].
8
Ag-NP Nanocomposite Cytotoxicity Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All animal experiments were conducted in accordance with policies of the Institutional Animal Care and Use Committee (IACUC) of National Taiwan University College of Medicine (NTUCM). The protocols used in this study were approved by IACUC (approved no. 20130364). ICR male and female mice (8 weeks old and 25 g weight) were acquired from Laboratory Animal Center (LAC), NTUCM. All mice were acclimatized for 1 week before being fed the solutions of Ag-NP nanocomposites at the dose of 5000 mg/kg body weight. In NC group, the mice were fed pure water while the PC group mice were injected intraperitoneally (i.p.) with mitomycin C (Sigma-Aldrich, USA) at dose of 1 mg/kg. Peripheral-blood cells were collected over the periods of 24h, 48h, and 72h. Micronucleus formations in the polychromatic erythrocytes were counted under microscope by Giemsa staining (Giemsa Stain Kit, Abcam, USA).
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!