To evaluate the morphological alterations induced by organotin(IV) complexes, MCF-7 and MDA-MB231 cell lines were cultured on a tissue culture flask with or without different treatments for 48 h. After incubation, cells were washed two times with PBS, fixed with methanol 100%, and stained with Wright-Giemsa (Sigma-Aldrich, WG32-1L). The culture flasks were examined under an Olympus IX71 Inverted Fluorescence Phase Contrast Microscope. The images were recorded with a Nikon Coolpix 4300 digital camera using the 20x objectives and analyzed by ImageJ software [32 (link)].
Coolpix 4300
Manufactured by Nikon
Sourced in Japan
The Nikon Coolpix 4300 is a digital camera with a 4.0-megapixel CCD image sensor. It features a 3x optical zoom lens, a 1.8-inch LCD display, and can capture images in JPEG format. The camera supports various shooting modes and has a built-in flash.
Automatically generated - may contain errors
Lab products found in correlation
Wright giemsa, by Merck Group (2 mentions)
Ix71 inverted fluorescence phase contrast microscope, by Olympus (2 mentions)
Ncl ki67p, by Leica (2 mentions)
Microphot fxa microscope, by Nikon (2 mentions)
Withalexafluor 488 and 594 igg secondary antibodies, by Thermo Fisher Scientific (2 mentions)
Nikoneclipse 90i fluorescence microscope, by Adobe (2 mentions)
Eclipse ts100, by Nikon (1 mentions)
Anti hpa1 c 20, by Santa Cruz Biotechnology (1 mentions)
Anti hpa2 c 17, by Santa Cruz Biotechnology (1 mentions)
Neutral red, by Merck Group (1 mentions)
Rhodamine b, by Merck Group (1 mentions)
Ab66039, by Abcam (1 mentions)
Immuno blot pvdf, by Bio-Rad (1 mentions)
Trans blot sd semi dry electrophoretic transfer cell, by Bio-Rad (1 mentions)
10 protocols using coolpix 4300
1
Morphological Alterations by Organotin Complexes
2
Quantitative Heparanase Immunostaining Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Heparanase immunostaining was performed using anti-HPA1 C-20 and anti-HPA2 C-17 antibodies (Santa Cruz Biotechnology®, Santa Cruz, CA, USA). A biotin-avidin-peroxidase complex was used to develop the reaction with 3,3′-diaminobenzidine used as the chromogen. Two independent observers scored 300 cells/slide as positive or negative according to the presence of staining for each of the above mentioned antibodies. The immunocytochemistry staining was analyzed by digital quantification. The slides were examined under a light microscope (Nikon Eclipse® TS100) to identify the areas that best represented typical immunostaining (hot spots). In each case, the quantitation was performed by digital analysis, and the values are expressed as the expression index (EI), following the methodology described by Matos and colleagues [33 (link)]. Photomicrographs of 640 × 480 pixels were obtained from consecutive nonoverlapping fields at 400× magnification with a digital camera (Nikon Coolpix® 4300) using the same parameters. The images were analyzed by a processing system and image analysis was performed using ImageLab (Softium Informática®, São Paulo, Brazil) using a micrometer scale (µm).
3
Immunohistochemical Analysis of MTH1 and Ki67
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunostaining, tissues were fixed overnight in formalin and stained
for human MTH1 (1/150 dilution; Novus Biologicals) and pan-proliferation marker
Ki67 (1/500 dilution; NCL-Ki67p, Leica Microsystems) as described
previously53 (link). Images
were photographed using a Nikon Microphot-FXA microscope and a Nikon Coolpix
4300 digital camera. For quantification, three random high-powered (200X) fields
(hpf) from each section were counted for a minimum of five different tumors per
sample (four for H23 shMTH1 to include all tumors formed at sites of injection).
Double immunofluorescent staining (DIF) was carried out using the MTH1 (1/150
dilution) and KRAS (1/100 dilution) antibodies, followed by incubation with
AlexaFluor 488 and 594 IgG secondary antibodies (1/500 dilution; Invitrogen) and
DAPI mounting medium (Vector Laboratories). Images were captured using a Nikon
Eclipse 90i fluorescence microscope and merged using Adobe Photoshop.
for human MTH1 (1/150 dilution; Novus Biologicals) and pan-proliferation marker
Ki67 (1/500 dilution; NCL-Ki67p, Leica Microsystems) as described
previously53 (link). Images
were photographed using a Nikon Microphot-FXA microscope and a Nikon Coolpix
4300 digital camera. For quantification, three random high-powered (200X) fields
(hpf) from each section were counted for a minimum of five different tumors per
sample (four for H23 shMTH1 to include all tumors formed at sites of injection).
Double immunofluorescent staining (DIF) was carried out using the MTH1 (1/150
dilution) and KRAS (1/100 dilution) antibodies, followed by incubation with
AlexaFluor 488 and 594 IgG secondary antibodies (1/500 dilution; Invitrogen) and
DAPI mounting medium (Vector Laboratories). Images were captured using a Nikon
Eclipse 90i fluorescence microscope and merged using Adobe Photoshop.
4
Assessing Mosquito Egg Follicle Defects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The assay has an advantage in that it can quickly assess whether follicles within the ovaries may contain defective eggshell prior to oviposition. Individual follicles of untreated, RNAi-Fluc, or RNAi-EOF1 mosquitoes at 96 h PBM were dissected and gently separated from the ovaries and transferred to glass scintillation vials. Rhodamine B (final concentration of 1 mM in H2O, Sigma-Aldrich) and neutral red (0.5%, Sigma-Aldrich) were used to stain primary follicles for 10 min on a rocking shaker and thoroughly rinsed with H2O. The stained primary follicles were photographed with a Coolpix 4300 (Nikon).
5
Immunoblotting Analysis of Cobalamin-Dependent Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Worms grown in the absence of CN-Cbl or in the presence of CN-Cbl or the CN-Cbl dodecylamine derivative were homogenized in 100 mmol/L potassium phosphate buffer (pH 7.0) at 4 °C. Each homogenate was centrifuged at 15,000×g for 10 min and the supernatant fraction was analyzed. We used a precast slab gel (PAGEL, type NPG-520L, ATTO Corporation, Tokyo, Japan) for electrophoresis of samples through a 5–20% (w/w) linear gradient of polyacrylamide in the presence of SDS. After electrophoresis, proteins were transferred to a PVDF membrane (Immuno-Blot PVDF, Bio-Rad Laboratories Inc., Hercules, CA, USA) in a Trans-Blot SD semi-dry electrophoretic transfer cell (Bio-Rad). The PVDF membrane was probed with an anti-MS antibody (ab66039, abcam®, Cambridge, MA, USA). We performed the immunodetection reactions using an anti-mouse IgG antibody secondary antibody (Promega KK, Tokyo, Japan) coupled to horseradish peroxidase and an immunoblot-staining kit for peroxidase (EzWestBlue, ATTO), according to the manufacturer’s instructions. A Protein Ladder One Triple-color kit (Nacalai Tesque Inc., Kyoto, Japan) was used to determine molecular mass. After the treated PVDF membrane was photographed using a digital camera (Coolpix 4300, Nikon, Japan), the intensities of the protein bands were calculated of Image J software [24] .
6
Immunohistochemical Analysis of MTH1 and Ki67
7
Morphological Alterations by Organotin Complexes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To evaluate the morphological alterations induced by organotin(IV) complexes, MCF-7 and MDA-MB231 cell lines were cultured on a tissue culture flask with or without different treatments for 48 h. After incubation, cells were washed two times with PBS, fixed with methanol 100%, and stained with Wright-Giemsa (Sigma-Aldrich, WG32-1L). The culture flasks were examined under an Olympus IX71 Inverted Fluorescence Phase Contrast Microscope. The images were recorded with a Nikon Coolpix 4300 digital camera using the 20x objectives and analyzed by ImageJ software [32 (link)].
8
Fabric Coating Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Optical photos of the fabrics coated with PCs were taken with a Nikon CoolPix4300 digital camera. The pictures were acquired under natural light, at the same time and environmental conditions, perpendicularly to the fabrics and at the distance of 15 cm. In the case of the multi-angle photographs the camera was disposed at different angles (0°, 45°and 90°) using a goniometer maintaining the same distance and using the same sample.
9
Quantifying Skin Flap Survival Rates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
On the 3rd and 7th days after surgery, the total flap and surviving skin areas were measured. The percentage survival rate of the flap was calculated using the following equation: Percentage of skin flap survival = survived skin area/total flap area × 10021 (link).
Flap survivability was assessed according to skin colour, eschar formation, and capillary refill5 (link). The desired skin surface areas were measured using ImageJ software after photography and calibration of the rat using a Nikon Coolpix 4300 digital camera (Nikon Corp., Tokyo, Japan)19 (link).
Flap survivability was assessed according to skin colour, eschar formation, and capillary refill5 (link). The desired skin surface areas were measured using ImageJ software after photography and calibration of the rat using a Nikon Coolpix 4300 digital camera (Nikon Corp., Tokyo, Japan)19 (link).
10
Immunohistochemical Analysis of Extracellular Matrix
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The detailed procedures for the immunohistochemical analysis have been previously published.11 (link) Briefly, the immunohistochemistry was performed to analyze protein immunolabeling for metalloproteinases, perlecan, and HPSE in 3 µm-thick histological slices. The primary antibodies used were anti-MMP-2 8B4 (sc-13595), anti-MMP-9 C-120 (sc-6840), anti-TIMP-2 H-140 (sc-5539), anti-perlecan H-300 (sc-25848), and anti-HPSE: HPA1 H-80 (sc-25825) (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) diluted in bovine serum albumin (BSA) in a ratio of 1:300. The slides were analyzed using a Nikon Eclipse TS100 optic microscope (Nikon Instruments, Melville, NY, USA) using the same light intensity and condenser height for all slides. The areas that best represented the immunolabeling of the slide were chosen and analyzed at a 400x magnification. Photomicrographs (640 × 480 pixels) of consecutive, nonoverlapping fields were obtained using a Nikon Coolpix 4300 (Nikon Corporation, Tokyo, Japan) digital camera with maximum optical zoom. Immunohistochemical labeling was quantified using the Scion ImageLab for Windows software (Scion Corporation, Frederick, MD, 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!