Biology subjects

Bradshaw, L.

Publications and source records attributed to Bradshaw, L..

2 recordsLinked to original sources

bioRxiv2026

Serial Immunohistochemistry for High-Dimensional Single-Cell Spatial Analysis of Human Kidney Biopsies

BackgroundTraditional immunohistochemistry (IHC) with chromogen detection has limited multiplex capacity, detecting at most 4 protein markers per tissue section simultaneously, thereby restricting comprehensive spatial analysis of valuable human biopsies. We developed and validated a robust serial IHC (sIHC) staining method to detect multiple antigens on a single kidney biopsy slide, maximizing data yield for diagnosing and studying complex kidney diseases. MethodsFormalin-fixed, paraffin-embedded kidney biopsy sections were subjected to repeated IHC/imaging cycles with antibody removal using an optimized sodium dodecyl sulfate-glycerol buffer stripping protocol. Images were then co-registered, and analysis was performed using a variety of methodologies, including color deconvolution, cell segmentation, and spatial clustering. ResultsThis optimized sIHC method successfully detected up to 20 antigens on a single slide. Combining image analysis and artificial intelligence software, for example with HALO (Indica Labs), the assay assembles high-dimensional images and enables quantitative histology and single-cell spatial analysis. Using this advanced method, we were able to identify rare cell populations, such as double-negative T cells, that are challenging to detect conventionally. ConclusionWe have developed a validated, high-capacity sIHC protocol that uses standard IHC procedures with commercially available, clinically validated off-the-shelf antibodies. This method is a valuable, cost-effective tool for obtaining extensive, high-dimensional single-cell-resolved spatial data from limited pathology samples, such as a human kidney biopsy.

pathology↗

bioRxiv2026

Adaptation to Environmental Variability Shapes Dormancy in Daphnia

Dormancy is a widespread adaptive strategy that allows organisms to survive in temporally varying habitats by suspending development and reproduction. Although environmental variability is expected to shape dormancy strategies, it remains unclear how differences in environmental variability and predictability influence both the production of dormant embryos and the termination of dormancy. We addressed these questions by comparing D. pulex and D. obtusa, two closely related species that inhabit environments differing in variability and predictability. We hypothesized that D. obtusa, which inhabits ephemeral environments, would exhibit a greater propensity for sexual reproduction and dormancy and would require stronger cues to break dormancy than D. pulex, which occurs in more permanent, predictable habitats. Consistent with our hypothesis, D. obtusa lineages produced significantly more males and ephippia than D. pulex when reared under identical laboratory conditions, indicating greater investment in sexual reproduction and dormancy. Contrary to our hypothesis, we found no difference in responsiveness to cues between the two species. Across species, embryos broke dormancy and hatched most readily after experiencing changes in cold and light, even if not experienced at the same time. In contrast, desiccation reduced the propensity to break dormancy. Together, these results indicate that species occupying more ephemeral environments invest more heavily in the production of dormant offspring, but that the environmental cues regulating dormancy termination appear broadly similar between species. This pattern suggests that while investment in dormancy may evolve in response to environmental variability, the mechanisms controlling dormancy termination are more conserved.

evolutionary biology↗