New instrument and methods for fast, diagnostic-quality histology of un-embedded bone marrow and lymph node specimens

Project Summary Overbone marrow biopsies are performed every year in the United Stateswhile hundreds of thousands more lymph node biospies are performedHistological evaluation of these biopsies is a critical component of care for hematologic diseases including leukemialymphomamyelodysplastic syndromemyeloproliferative disease and non neoplastic conditions such as viral infections and autoimmune conditionsWe have developed a platform that we consider a paradigm shift in the histologic examination of tissuesIt is based on a new chemical processimagingand image processing approach that we have dubbed Clearing Histology with MultiPhoton microscopyCHiMPThe CHiMP technology enables visual analysis of entire intactun embedded and uncut specimens within a short time frame and with a resolution that is amenable to primary diagnosisThe significant clinical benefits includepotential for same day diagnosislabor and cost savingsaccess toD perspectiveincreased visual data from same specimencomplete tissue preservation for ancillary studies such as DNA analysis andinherent benefits of digital data such as reduced risk of lossready remote review by expertsand amenability to machine learning toolsHematopoietic tissue evaluation would similarly benefit from these advantagesbut unfortunately the systems developed thus far lack the resolution typically needed for visual examination of hematopoietic tissuesFor this Phase I SBIR proposalan objective is to develop customized optics to improve the resolution of our current microscopes and thereby enable use in the specialized field of hematologyCommercially available objective lenses that are compatible with our immersion medium are either limited to numerical aperturesNAthat are less than oneaffecting resolution and image qualityor have insufficient working distances for imaging past the coverslip and surface roughness to obtain complete sectionsWe will design and test a custom objective lens with high NA and long working distancesuited for our proprietary reagentsIntegrating such a lens into our microscope will also require the design of a custom scan lenscustom beam conditioning opticsand a custom polygon scannerAn associated goal is to develop a novel approach to preparing bone marrow aspiration specimens that will make them amenable to imaging with CHiMPpotentially reducing the need for core biopsies by permitting unambiguous morphologic categorization of cell subtypes in their architectural contextwithout the routine need for immunohistochemistryand while preserving nucleic acids for molecular genetic evaluation Project Narrative Applikate Technologies has developed a powerful platform for histological evaluation of tissue called Clearing Histology with MultiPhoton Microscopyor CHiMPThis platform has many advantages over traditional approachesincluding same day turn aroundreduced labor costspreservation of tissue for DNA analysisand direct to digital imaging for ease of consultation with remote expertsThis proposal seeks to develop custom optics to enable very high resolution imaging of bone marrow and lymph node samples that are critical for diagnosing diseases such as leukemia and lymphoma