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Photomicrography

Photomicrography is the art and science of capturing high-quality images of microscopic specimens.
This techneque allows researchers to visualize and analyze the detailed structures of cells, tissues, and other biological samples with great precision.
Photomicrography involves the use of specialized cameras, microscopes, and imaging software to produce clear, high-resolution micrographs that can be used for scientific study, medical diagnosis, and educational purposes.
Effective photomicrography requires a thorough understanding of opticsl principles, camera settings, and specimen preparation methods to optimize image quality and reproducibility.
Leveraging the latest advancements in digital imaging and AI-powered analysis can further enhance the accuracy and efficiency of photomicrography research.

Most cited protocols related to «Photomicrography»

1
Retinal Histology Database and Imaging
Cited 38 times
Original literature from 1900 until 2009 describing the histology of the retina were reviewed to find relevant information for a database of representative values (Table 1). Repetitive or derivative works were not used. This assembled information, along with the photomicrographs in the publications, was used to create drawings of the outer retina using CorelDRAW X5 (Corel Corporation, Ottawa, Canada). The drawings were made to scale, with equal horizontal and vertical scaling. Three representations were made of photoreceptors: cones from the central fovea and rods and a cone from the perifoveal region (as per Polyak,35 2 mm temporal to the fovea). In the drawing, junctional complexes between the Muller cells and the photoreceptors, which in aggregate form the ELM, were labeled as the ELM. Similarly the junctional complexes girdling the RPE cells were labeled “Verhoeff membrane” to be consistent with past histologic nomenclature.
Drawings were compared with a representative scan obtained of one of us (R.F.S.) using the Heidelberg Spectralis HRA+OCT (Heidelberg Engineering, Vista, CA) in high-resolution mode. This instrument has an axial resolution of approximately 7 μm. A longitudinal reflectance profile (LRP) from a horizontal scan taken through the fovea center was created by averaging pixels across at each level of 10 adjacent A-scans using ImageJ (Version 1.44f; National Institutes of Health, Bethesda, MA). Additional scans were obtained of other subjects that did not differ from what is shown in this report and scans obtained with the Cirrus HD-OCT (Carl Zeiss Meditec, Dublin, CA), which also showed similar results. Therefore, only one scan will be shown in the comparison to the model. The LRPs were determined from the display information of the Heidelberg Spectralis.
SPAIDE R.F, & CURCIO C.A. (2011). ANATOMICAL CORRELATES TO THE BANDS SEEN IN THE OUTER RETINA BY OPTICAL COHERENCE TOMOGRAPHY: Literature Review and Model. Retina (Philadelphia, Pa.), 31(8), 1609-1619.
Publication 2011
Cells Intercellular Junctions Photomicrography Photoreceptor Cells Radionuclide Imaging Retina Retinal Cone Rod Photoreceptors Tissue, Membrane
2
Methodology for Rat Brain Atlas Production
Cited 30 times
Detailed methodology for producing the rat brain atlas is provided in the first three editions (Swanson, 1992, 1998, 2004) that are available as open access legacy resources (Swanson, 2015b) at larrywswanson.com. Briefly, after many attempts (starting in 1974) to obtain a complete series of transverse histological sections suitable for an atlas, one was obtained in 1982 from a 315‐g adult male Sprague‐Dawley rat that had been perfused with 4% paraformaldehyde and embedded in celloidin to hold separate parts of sections in place during mounting. All procedures for rats complied with NIH and institutional guidelines current from 1974 to 1982; the work on the atlas brain was done at the Salk Institute for Biological Studies, La Jolla, CA. Every section through the brain was collected, stained, and mounted; the first 133 sections through the olfactory bulbs were 30 µm thick, whereas the last 423 sections through the rest of the brain were 40 µm thick. The sections were stained with thionin and covered with DPX.
Because celloidin‐embedded tissue shrinks considerably and differentially in the rostro‐caudal, medio‐lateral, and dorso‐ventral dimensions, two Cartesian coordinate systems for the sections were produced. The first is a strictly physical coordinate system, corresponding to dimensions in the tissue sections themselves. The second is a stereotaxic coordinate system that ideally would be based on the dimensions of the brain within the skull of the intact, living animal. Fortunately, this brain was cut in virtually the same transverse plane as the stereotaxic rat brain atlas of Paxinos and Watson (1986), based on unembedded, frozen‐sectioned brains that suffered very little shrinkage. Because researchers have found the stereotaxic coordinates in Paxinos and Watson (1986) to be the best available, they were adopted for our brain as the second set of coordinates.
Photomicrographs of selected histological sections were obtained by placing the sections in an Omega enlarger with a point light source, projecting an image of the section onto a 4 × 5 inch sheet of Kodak Kodalith Ortho (2556) film, developing the film in Kodak Kodalith fine line developer, and printing with a Durst enlarger and Schneider Kreuzanch Componon‐S lens (f/150 mm) on 11 × 14 inch sheets of Kodak Kodabrome II RC paper, contrast grade F5. After 35 years, these thick celloidin sections are unsuitable for high resolution digital scanning because they are not completely flat and because the DPX has retracted in places, creating random “bubbles” of air between tissue section and coverslip. However, most areas of the sections remain suitable for microscopic examination.
, & Swanson L.W. (2018). Brain maps 4.0—Structure of the rat brain: An open access atlas with global nervous system nomenclature ontology and flatmaps. The Journal of Comparative Neurology, 526(6), 935-943.
Publication 2018
Adult Animals Biopharmaceuticals Brain Celloidin Cranium Lens, Crystalline Light Males Microscopy Olfactory Bulb paraform Photomicrography Physical Examination Rats, Sprague-Dawley Thionins Tissues
3
Eosinophilic Esophagitis Diagnostic Markers
Cited 26 times

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Publication 2009
Biopsy Diagnosis Eosinophil Esophagus Immunohistochemistry Medical Laboratory Personnel Patients Photomicrography Stomach Tissues
4
Quantifying Microglial Morphology and Complexity
Cited 25 times
A ramified cell is one that has a complicated network of processes that originate from the cell soma. A change in microglia ramification indicates a microglial response to an altered physiologic status, in this case induced by mFPI. ImageJ software (National Institute of Health, https://imagej.nih.gov/ij/34 (link)) and appropriate plugins (i.e. FFT bandpass filter, unsharp mask and close) were consistently used prior to converting all photomicrographs to binary and skeletonized images. In addition to creating skeletonized images, cell somas were manually counted for each photomicrograph. The Analyze Skeleton Plugin (developed by and maintained here: http://imagej.net/AnalyzeSkeleton31) was then applied to the skeleton image which tags skeletal features relevant to microglia ramification: slab voxels (orange, process length) and endpoints (blue). Figure 2 illustrates the workflow process to convert an entire original photomicrograph to a plugin tagged image (original, binary and skeleton); cropped images and an overlay of skeleton to original image is provided for detail and to illustrate that skeletons are representative of the original image. We summarized the number of process endpoints and length from the Analyze Skeleton plugin data output and normalized all data by the number of microglia cell somas in each image to calculate the number of microglia endpoints/cell and microglia process length/cell. In Table 1, we summarize skeleton analysis measures (endpoints and process length/cell) in terms of measure, unit, range, scale, and interpretation.

Skeleton analysis of microglia morphologies in Iba1 stained tissue. (a) The process to prepare photomicrographs for skeleton analysis. Original photomicrographs were subjected to a series of uniform ImageJ plugin protocols prior to conversion to binary images; binary images were then skeletonized. An overlay of a resulting skeletonized image (in green) and original photomicrograph shows the relationship between skeleton and photomicrograph. Cropped photomicrographs (below) show additional detail and all skeleton analysis was completed on full sized photomicrographs (Scale bar = 10 µm). (b) The skeletonized images are processed using the Analyze Skeleton plugin (maintained here: http://imagej.net/AnalyzeSkeleton) to identify and tag skeletonized processes as orange, endpoints as blue, and junctions as purple. The tagged data are then organized and data output summarized.

Summary of microglia morphology measures.

MeasureUnitRangeScaleSamplingInterpretation
Process length22 Summedµm/cellContinuousPhotomicro-graph6 photomicrographs/animalCell ramification
Process endpoints22 Summed#/cellContinuousPhotomicro-graph6 photomicrographs/animalCell ramification
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\begin{document}$$\mathrm{regression}\,\mathrm{slope}[\frac{In(N)}{In(\varepsilon )}]$$\end{document}regressionslope[In(N)In(ε)]		DB1-2Individual cell24 cells/animalCell complexity
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\begin{document}$$\frac{\#\,\mathrm{of}\,\mathrm{pixels}\,\mathrm{within}\,\mathrm{cell}\,\mathrm{outline}}{\mathrm{area}\,\mathrm{of}\,\mathrm{convex}\,\mathrm{hull}}$$\end{document}#ofpixelswithincelloutlineareaofconvexhull		\documentclass[12pt]{minimal}
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Morrison H., Young K., Qureshi M., Rowe R.K, & Lifshitz J. (2017). Quantitative microglia analyses reveal diverse morphologic responses in the rat cortex after diffuse brain injury. Scientific Reports, 7, 13211.
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Publication 2017
Cell Body Cells Microglia Photomicrography physiology Skeleton Tissues
5
Prostate stem cell lineage tracing
Cited 25 times
The Nkx3.1CreERT2/+ allele was generated by gene targeting using standard techniques; the Nkx3.1 null mutant mice have been previously described21 (link). R26R-lacZ and Pten conditional mutant mice were obtained from the Jackson Laboratory Induced Mutant Resource; the R26R-YFP mice were provided by Dr. Frank Costantini. All lines were maintained on a hybrid C57BL/6-129/Sv strain background.
Castration of adult male mice was performed using standard techniques. For tamoxifen induction of Cre activity in mice containing Nkx3.1CreERT2/+, mice were administered 9 mg/40 g tamoxifen for 4 consecutive days. For prostate regeneration, physiological levels of testosterone (1.875 µg/hr) were administered for four weeks by subcutaneous implantation of mini-osmotic pumps (Alzet)45 (link). When included, BrdU (100 mg/kg) was administered once daily during the first three days of regeneration. For single-cell transplantation, single YFP+ cells were isolated by mouth-pipetting under epifluorescence illumination from a dissociated prostate cell suspension obtained from castrated and tamoxifen-induced Nkx3.1CreERT2/+; R26R-YFP/+ mice. A single YFP+ cell (or YFP− cell as a control) was recombined with 2.5 × 105 rat urogenital sinus mesenchyme cells in a 10 µl collagen pad, followed by transplantation under the kidney capsule of nude mice and harvesting after 10–12 weeks.
Cryosections were stained with primary antibodies as listed in Supp. Table 5, and counterstained with TOPRO3 or DAPI (Invitrogen/Molecular Probes). Secondary antibodies were labeled with Alexa Fluor 488 , 555, or 594 (Invitrogen/Molecular Probes). Immunofluorescence staining was imaged using a Leica TCS5 spectral confocal microscope. Cell counting was performed manually using confocal photomicrographs with at least three animals for each experiment or genotype analyzed.
Wang X., Kruithof-de Julio M., Economides K.D., Walker D., Yu H., Halili M.V., Hu Y.P., Price S.M., Abate-Shen C, & Shen M.M. (2009). A luminal epithelial stem cell that is a cell of origin for prostate cancer. Nature, 461(7263), 495-500.
Publication 2009
Adult alexa fluor 488 Alleles Animals Antibodies Bromodeoxyuridine Capsule Cells Cell Transplantation Collagen Cryoultramicrotomy DAPI Fluorescent Antibody Technique Genotype Hybrids Kidney LacZ Genes Light Male Castration Mesenchyma Mice, Knockout Mice, Nude Microscopy, Confocal Molecular Probes Mus Oral Cavity Orchiectomy Osmosis Ovum Implantation Photomicrography physiology Prostate PTEN protein, human Regeneration Sinuses, Nasal Strains System, Genitourinary Tamoxifen Testosterone Transplantation

Most recents protocols related to «Photomicrography»

1
Quantifying Marrow Bone in Femur Sections
The paraformaldehyde fixed femur samples were decalcified (6 weeks; in 10% EDTA phosphate buffer solution), dehydrated (in ethyl alcohol), hyalinized (in xylene), and the proximal diaphysis was embedded in paraffin wax. Sample sections (5 μm thick; mounted on glass slides) were stained with toluidine blue and scanned using an optical microscope (BX46; Olympus, Japan). Three photomicrographs (40× magnification) were taken from each sample. The area percentage of MB is expressed as the percentage of MB in the region of the marrow cavity where the MB is located (ImageJ 1.8.0. software).
Yan J., Wang J., Chen J., Shi H., Liao X., Pan C., Liu Y., Yang X., Ren Z, & Yang X. (2023). Adjusting phosphate feeding regimen according to daily rhythm increases eggshell quality via enhancing medullary bone remodeling in laying hens. Journal of Animal Science and Biotechnology, 14, 17.
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Publication 2023
Buffers Dental Caries Diaphyses Edetic Acid Ethanol Femur Light Microscopy Marrow Paraffin paraform Phosphates Photomicrography Tolonium Chloride Xylene
2
Histological Analysis of Iron Deposition in COPD Lung Tissue
The study was reviewed and approved by the Duke University Institutional Review Board (Durham, NC). An existing cohort of patients diagnosed to have chronic obstructive pulmonary disease (COPD; some combination of chronic bronchitis and emphysema) was searched and six individuals identified. Blocks of lung tissue collected at autopsy were retrieved from archives.
Perls’ Prussian blue was employed to stain iron. Hale’s stain was used as an assay for in situ iron binding capacity20 (link). The background stain was nuclear fast red. Tissue was stained for an iron importer and storage protein. Five micron tissue sections were cut, floated on a protein-free water bath, mounted on silane treated slides, and air-dried overnight. Sections were then deparaffinized and hydrated to 95% alcohol (xylene for 10 min, absolute alcohol for 5 min, and 95% alcohol for 5 min). Endogenous peroxidase activity was blocked with 0.6% H2O2 in absolute methanol for eight minutes. Slides were rinsed in 95% alcohol for 2 min, placed in deionized H2O, and washed in PBS. After treatment with Cyto Q Background Buster (Innovex Biosciences, Richmond, CA) for 10 min, slides were incubated with the primary antibody diluted in 1% bovine serum albumin for 45 min at 37 °C in PBS. Primary antibodies used in this investigation were to divalent metal transport 1 (DMT1) (generously provided by Dr. Funmei Yang of the University of Texas, San Antonio, TX) used at a dilution of 1:200 and ferritin (Dako, Carpinteria, CA) used at a dilution of 1:200. Slides were incubated with biotinylated linking antibody from Stat-Q Staining System (Innovex Biosciences) for ten minutes at room temperature, washed with PBS, and peroxidase enzyme label from Stat-Q Staining System (Innovex Biosciences) applied. After incubation for ten minutes at room temperature and washes with PBS, tissue sections were developed with 3,3′diaminobenzidine-tetrahydrochloride for three minutes at room temperature. Sections were counterstained with hematoxylin, dehydrated through alcohols, cleared in xylene and coverslipped using a permanent mounting media. Photomicrographs were obtained using a Nikon Eclipse E600 microscope (Tokyo, Japan) with 10×/40× objective lens coupled with QCapture software (QImaging, Surrey, British Columbia, Canada).
Ghio A.J., Soukup J.M., Dailey L.A, & Roggli V.L. (2023). Mucus increases cell iron uptake to impact the release of pro-inflammatory mediators after particle exposure. Scientific Reports, 13, 3925.
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Publication 2023
Absolute Alcohol Aftercare Antibodies Apnea Autopsy Bath Biological Assay Bronchitis, Chronic Chronic Obstructive Airway Disease E-600 Ethanol Ethics Committees, Research ferric ferrocyanide Ferritin Glycogen Branching Enzyme Hematoxylin Immunoglobulins Iron Lens, Crystalline Metals Methanol Microscopy Patients Peroxidase Peroxide, Hydrogen Photomicrography Proteins Pulmonary Emphysema Serum Albumin, Bovine Silanes Staphylococcal Protein A Technique, Dilution Tissues Xylene
3
Histological Analysis of Rat Hippocampus
The isolated hippocampus of the brains (6 rats for each group) was fixed in 10%
formalin, embedded in paraffin wax, sectioned, and stained. The sections were
examined under a light microscope, and photomicrographs were done to assess
histopathological changes.
El-din Hussein A.S., Abou-El Nour R.K., Khorshid O.A, & Osman A.S. (2023). Study of the possible effect of sacubitril/valsartan combination versus valsartan on the cognitive function in Alzheimer’s disease model in rats. International Journal of Immunopathology and Pharmacology, 37, 03946320231161469.
Publication 2023
Brain Formalin Light Microscopy Paraffin Photomicrography Rattus norvegicus Seahorses
4
Histological Analysis of Retinal Tissue

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Publication 2023
Ethanol Eye Light Microscopy Paraffin Embedding paraform Photomicrography Retina Xylene
5
Neuroanatomical Volume Measurements
The volumes the DG, GC layer and hippocampus proper for each animal was measured in 6 serial sections on photomicrographs acquired using a Leica ICC50 HD digital camera using a free hand tool of ImageJ plugin, VOLUMEST [32 (link), 33 (link)].
Xhakaza N.K., Nkomozepi P, & Mbajiorgu E.F. (2023). Boophone disticha attenuates five day repeated forced swim-induced stress and adult hippocampal neurogenesis impairment in male Balb/c mice. Anatomy & Cell Biology, 56(1), 69-85.
Publication 2023
Animals Fingers Photomicrography Seahorses

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1
Bx51 microscope by Olympus
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2
Bx41 microscope by Olympus
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Eclipse 80i by Nikon
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Axiovision software by Zeiss
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AxioVision is a software package developed by ZEISS. It provides a user interface for the control and operation of ZEISS microscopes and imaging systems. The software enables the capture, processing, and analysis of digital images and data generated by ZEISS hardware.
6
Axiophot microscope by Zeiss
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More about "Photomicrography"

Photomicrography: The Art and Science of Microscopic Imaging

Phtotomicrography, also known as microscopic imaging or microimaging, is the technique of capturing high-quality, detailed images of microscopic specimens.
This powerful tool allows researchers, scientists, and medical professionals to visualize and analyze the intricate structures of cells, tissues, and other biological samples with unparalleled precision.
The photomicrography process involves the use of specialized cameras, microscopes, and imaging software to produce clear, high-resolution micrographs that can be utilized for scientific study, medical diagnosis, and educational purposes.
Effective photomicrography requires a deep understanding of optical principles, camera settings, and specimen preparation methods to optimize image quality and reproducibility.
Advanced technologies, such as digital imaging and AI-powered analysis, have further enhanced the accuracy and efficiency of photomicrography research.
Tools like the BX51 and BX41 microscopes, Image-Pro Plus 6.0, Eclipse 80i, and AxioVision software provide researchers with powerful imaging capabilities, while the AxioCam, AxioVision 4.8, and DAPI staining technique offer additional ways to capture and analyze microscopic data.
By leveraging the latest advancements in photomicrography, researchers can take their microscopic imaging to new heights, unlocking a wealth of insights and discoveries that can drive scientific progress and improve medical outcomes.
Whether you're a seasoned professional or a curious learner, the world of photomicrography offers endless opportunities for exploration and innovation.