Protocol for Three-dimensional Confocal Morphometric Analysis of Astrocytes

Bagheri, Maryam; Rezakhani, Arjang; Roghani, Mehrdad; Joghataei, Mohammad Taghi; Mohseni, Simin

doi:10.3791/53113-v

Cited by 2 publications

(2 citation statements)

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“…Elevated serum GFAP is, therefore, a biomarker for neurotoxicity, specifically injury to neuronal and astroglial cells (Yang and Wang, 2015). Variations in GFAP levels can manifest in response to traumatic injuries, neurodegenerative or neuroinflammatory processes (Bagheri et al, 2015, Abdelhak et al, 2019), neuropathology (Wang and Parpura, 2016), disruption of the blood-brain barrier (Torres-Platas et al, 2016), and brain vascular inflammation. Serum and cerebrospinal fluid NSE elevation is an effective biomarker for neuronal loss and impairment, brain injury, and neuroinflammation (Park et al, 2019, Bagheri et al, 2022, Haque et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

In children with transfusion-dependent thalassemia, inflammation and neuronal damage biomarkers are associated with affective and chronic fatigue symptoms

Ridhaa,

Al-Hakeim,

Kahlol

et al. 2023

Preprint

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BackgroundPatients with transfusion-dependent thalassemia (TDT) are vulnerable to neurotoxicity due to frequent blood transfusions and the subsequent iron overload (IO) and inflammation. As a result, affective (depression and anxiety) and chronic fatigue syndrome (CFS) symptoms may develop.AimsTo investigate the potential association between TDT and neuronal injury, as assessed with serum concentrations of neuronal damage biomarkers, including neurofilament light (NFL), glial fibrillary acidic protein (GFAP), neuron-specific enolase (NSE), and nestin.MethodsWe investigated the associations between those CNS injury biomarkers, neuro-immune markers (C-reactive protein (CRP), interleukin (IL)-6, and IL-10), calcium, magnesium, copper and zinc, and the Fibro-Fatigue (FF), the Children’s Depression Inventory (CDI), and the Spence Children’s Anxiety Scale (SCAS) scores in 126 children with TDT and 41 healthy children.ResultsTDT children show significant increases in IO, FF, CDI, and SCAS scores, serum NSE, GFAP, NF-L, CRP, copper, IL-6, and IL-10, and lowered magnesium, zinc, and calcium as compared with healthy children. There were significant correlations between the CDI score and NFL, NSE and GFAP; SCAS score and NFL, and FF score and NFL and GFAP. The neuronal damage biomarkers (except nestin) were significantly associated with inflammatory, erythron (hematocrit and hemoglobin) and IO (iron and ferritin) biomarkers.ConclusionsTDT is characterized by intertwined increases in neuronal injury biomarkers and neuropsychiatric symptoms suggesting that TDT-associated neurotoxicity plays a role in affective symptoms and CFS due to TDT. Inflammation and neurotoxicity are novel drug targets for the prevention of affective symptoms and CFS due to TDT.

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Section: Discussionmentioning

confidence: 99%

In children with transfusion-dependent thalassemia, inflammation and neuronal damage biomarkers are associated with affective and chronic fatigue symptoms

Ridhaa,

Al-Hakeim,

Kahlol

et al. 2023

Preprint

View full text Add to dashboard Cite

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“…Area (Williams et al, 2010), Cell coverage (Williams et al, 2010), Colocalization (Virgone-Carlotta et al, 2013, Count (Choi et al, 2010;Kelber et al, 2010;Elliott et al, 2015), Density (Virgone-Carlotta et al, 2013), Distance between cells (Williams et al, 2010), Length (Choi et al, 2010;Kelber et al, 2010;Virgone-Carlotta et al, 2013), Process (Branch/Dendrite) Count (Elliott et al, 2015), Process (Branch/Dendrite) intersections (Elliott et al, 2015) Area coverage (Williams et al, 2010;Naguib et al, 2012), Colocalization (Li et al, 2013), Count (Choi et al, 2010;Li et al, 2013;Bolea et al, 2014), Distance between cells (Williams et al, 2010), Intensity (Ghiretti and Paradis, 2011;Naguib et al, 2012), Length (Choi et al, 2010) Area (Bayerl et al, 2016), Axon length (Kemp et al, 2016), Cell density (Bayerl et al, 2016), Colocalization (Ugbode et al, 2014;Wang et al, 2017), Count (Bayerl et al, 2016;Kemp et al, 2016), Intensity (Bayerl et al, 2016;Chu et al, 2020), Size (Kemp et al, 2016), Surface Area (Bagheri et al, 2015), Volume (Bagheri et al, 2015;Bayerl et al, 2016;Chu et al, 2020) Image One limitation we noted is the MeSH term PubMed search left out many papers citing some of the more popular software. For example, the authors use image proce...…”

Section: Software Comparison Tablementioning

confidence: 99%

Prevalence and practices of immunofluorescent cell image processing: a systematic review

Helmbrecht

Lin

Janakiraman³

et al. 2023

Front. Cell. Neurosci.

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BackgroundWe performed a systematic review that identified at least 9,000 scientific papers on PubMed that include immunofluorescent images of cells from the central nervous system (CNS). These CNS papers contain tens of thousands of immunofluorescent neural images supporting the findings of over 50,000 associated researchers. While many existing reviews discuss different aspects of immunofluorescent microscopy, such as image acquisition and staining protocols, few papers discuss immunofluorescent imaging from an image-processing perspective. We analyzed the literature to determine the image processing methods that were commonly published alongside the associated CNS cell, microscopy technique, and animal model, and highlight gaps in image processing documentation and reporting in the CNS research field.MethodsWe completed a comprehensive search of PubMed publications using Medical Subject Headings (MeSH) terms and other general search terms for CNS cells and common fluorescent microscopy techniques. Publications were found on PubMed using a combination of column description terms and row description terms. We manually tagged the comma-separated values file (CSV) metadata of each publication with the following categories: animal or cell model, quantified features, threshold techniques, segmentation techniques, and image processing software.ResultsOf the almost 9,000 immunofluorescent imaging papers identified in our search, only 856 explicitly include image processing information. Moreover, hundreds of the 856 papers are missing thresholding, segmentation, and morphological feature details necessary for explainable, unbiased, and reproducible results. In our assessment of the literature, we visualized current image processing practices, compiled the image processing options from the top twelve software programs, and designed a road map to enhance image processing. We determined that thresholding and segmentation methods were often left out of publications and underreported or underutilized for quantifying CNS cell research.DiscussionLess than 10% of papers with immunofluorescent images include image processing in their methods. A few authors are implementing advanced methods in image analysis to quantify over 40 different CNS cell features, which can provide quantitative insights in CNS cell features that will advance CNS research. However, our review puts forward that image analysis methods will remain limited in rigor and reproducibility without more rigorous and detailed reporting of image processing methods.ConclusionImage processing is a critical part of CNS research that must be improved to increase scientific insight, explainability, reproducibility, and rigor.

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Protocol for Three-dimensional Confocal Morphometric Analysis of Astrocytes

Cited by 2 publications

References 0 publications

In children with transfusion-dependent thalassemia, inflammation and neuronal damage biomarkers are associated with affective and chronic fatigue symptoms

In children with transfusion-dependent thalassemia, inflammation and neuronal damage biomarkers are associated with affective and chronic fatigue symptoms

Prevalence and practices of immunofluorescent cell image processing: a systematic review

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