Wide field block face imaging using deep ultraviolet induced autofluorescence of the human brain

Karthik, Srinivasa; Joseph, Jayaraj; Jayakumar, Jaikishan; Manoj, Rahul; Shetty, Mahabalesh; Bota, Mihail; Verma, Richa; Mitra, Partha P.; Sivaprakasam, Mohanasankar

doi:10.1016/j.jneumeth.2023.109921

Cited by 3 publications

(2 citation statements)

References 32 publications

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“…Every two consecutive sections were transferred to two adjacent large format 6" × 8" slides for alternate series of staining with Nissl and Hematoxylin and Eosin (H&E) (data not shown). Block Face Imaging with ultraviolet and white light was taken before each section using an excitation and emission wavelength of 254 nm (UV-C) and 400-500 nm (visible spectrum), respectively (Karthik et al, 2023).…”

Section: Cryosectioning Using the Tape Transfer Techniquementioning

confidence: 99%

A technology platform for standardized cryoprotection and freezing of large-volume brain tissues for high-resolution histology

Kumarasami,

Verma,

Pandurangan

et al. 2023

Front. Neuroanat.

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Understanding and mapping the human connectome is a long-standing endeavor of neuroscience, yet the significant challenges associated with the large size of the human brain during cryosectioning remain unsolved. While smaller brains, such as rodents and marmosets, have been the focus of previous connectomics projects, the processing of the larger human brain requires significant technological advancements. This study addresses the problem of freezing large brains in aligned neuroanatomical coordinates with minimal tissue damage, facilitating large-scale distortion-free cryosectioning. We report the most effective and stable freezing technique utilizing an appropriate choice of cryoprotection and leveraging engineering tools such as brain master patterns, custom-designed molds, and a continuous temperature monitoring system. This standardized approach to freezing enables high-quality, distortion-free histology, allowing researchers worldwide to explore the complexities of the human brain at a cellular level. Our approach combines neuroscience and engineering technologies to address this long-standing challenge with limited resources, enhancing accessibility of large-scale scientific endeavors beyond developed countries, promoting diverse approaches, and fostering collaborations.

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Section: Cryosectioning Using the Tape Transfer Techniquementioning

confidence: 99%

A technology platform for standardized cryoprotection and freezing of large-volume brain tissues for high-resolution histology

Kumarasami,

Verma,

Pandurangan

et al. 2023

Front. Neuroanat.

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“…Here we apply our novel methodology, including postmortem inskull structural MRI and block-face imaging, of serial sections across the entire brain (Karthik et al, 2023) to supplement our detailed histological analyses of the cortical cytoarchitecture of the mesial surface in relation to the early sulcal roots, laminar organization of the developing cerebral cortex, and the pattern of development of radial glial cells in the cerebral cortex and subcortical regions in 13-15 GW human fetal brain.…”

Section: Introductionmentioning

confidence: 99%

Histological characterization and development of mesial surface sulci in the human brain at 13–15 gestational weeks through high‐resolution histology

Verma,

Jayakumar,

Folkerth

et al. 2024

J of Comparative Neurology

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Cellular‐level anatomical data from early fetal brain are sparse yet critical to the understanding of neurodevelopmental disorders. We characterize the organization of the human cerebral cortex between 13 and 15 gestational weeks using high‐resolution whole‐brain histological data sets complimented with multimodal imaging. We observed the heretofore underrecognized, reproducible presence of infolds on the mesial surface of the cerebral hemispheres. Of note at this stage, when most of the cerebrum is occupied by lateral ventricles and the corpus callosum is incompletely developed, we postulate that these mesial infolds represent the primordial stage of cingulate, callosal, and calcarine sulci, features of mesial cortical development. Our observations are based on the multimodal approach and further include histological three‐dimensional reconstruction that highlights the importance of the plane of sectioning. We describe the laminar organization of the developing cortical mantle, including these infolds from the marginal to ventricular zone, with Nissl, hematoxylin and eosin, and glial fibrillary acidic protein (GFAP) immunohistochemistry. Despite the absence of major sulci on the dorsal surface, the boundaries among the orbital, frontal, parietal, and occipital cortex were very well demarcated, primarily by the cytoarchitecture differences in the organization of the subplate (SP) and intermediate zone (IZ) in these locations. The parietal region has the thickest cortical plate (CP), SP, and IZ, whereas the orbital region shows the thinnest CP and reveals an extra cell‐sparse layer above the bilaminar SP. The subcortical structures show intensely GFAP‐immunolabeled soma, absent in the cerebral mantle. Our findings establish a normative neurodevelopment baseline at the early stage.

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A three-dimensional histological cell atlas of the developing human brain

Jayakumar,

Sivaprakasam,

Verma

et al. 2024

Preprint

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The human brain is believed to contain a full complement of neurons by the time of birth together with a substantial amount of the connectivity architecture, even though a significant amount of growth occurs postnatally. The developmental process leading to this outcome is not well understood in humans in comparison with model organisms. Previous magnetic resonance imaging (MRI) studies give three-dimensional coverage but not cellular resolution. In contrast, sparsely sampled histological or spatial omics analyses have provided cellular resolution but not dense whole brain coverage. To address the unmet need to provide a quantitative spatiotemporal map of developing human brain at cellular resolution, we leveraged tape-transfer assisted serial section histology to obtain contiguous histological series and unbiased imaging with dense coverage. Interleaved 20μ thick Nissl and H&E series and MRI volumes are co-registered into multimodal reference volumes with 60μ isotropic resolution, together with atlas annotations and a stereotactic coordinate system based on skull landmarks. The histological atlas volumes have significantly more contrast and texture than the MRI volumes. We computationally detect cells brain-wide to obtain quantitative characterization of the cytoarchitecture of the developing brain at 13-14 and 20-21 gestational weeks, providing the first comprehensive regional cell counts and characterizing the differential growth of the different brain compartments. Morphological characteristics permit segmentation of cell types from histology. We detected and quantified brain-wide distribution of mitotic figures representing dividing cells, providing an unprecedented spatiotemporal atlas of proliferative dynamics in the developing human brain. Further, we characterized the abundance and distribution of Cajal-Retzius cells, a transient cell population that plays essential roles in organizing glutamatergic cortical neurons into layers. Together, our study provides an unprecedented quantitative window into the developing human brain and the reference volumes and coordinate space should be useful for integrating spatial omics data sets with dense histological context.

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Wide field block face imaging using deep ultraviolet induced autofluorescence of the human brain

Cited by 3 publications

References 32 publications

A technology platform for standardized cryoprotection and freezing of large-volume brain tissues for high-resolution histology

A technology platform for standardized cryoprotection and freezing of large-volume brain tissues for high-resolution histology

Histological characterization and development of mesial surface sulci in the human brain at 13–15 gestational weeks through high‐resolution histology

A three-dimensional histological cell atlas of the developing human brain

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