RAPID: DEBI-t: development of a Deep Exploration Biosphere Investigative Tool

了解深层生命在地下生物圈中的分布是推进我们对地球上生命进化和生态系统理解的一个重大挑战。迄今为止，所有用于探测深层生物圈中微生物生命的方法都涉及对从钻孔中回收的材料进行非原位分析。通过提取细胞或细胞组分，或通过将染料应用于样品并进行细胞计数来分析材料的微生物生物量。后一种方法是行业中最标准的方法，并且代表了在显微镜下通过眼睛计数荧光细胞的非常耗时和劳动密集的动手操作。 目前的方法是非常费力和低效的过程，涉及细胞损失和可能对微生物生态学产生影响的矿物学背景信息的损失。为了应对与检测和量化天然基质中微生物生命相关的挑战，PI要求RAPID资助开发一种新型的原位井下测井工具，用于检测海底下钻孔中的微生物生命。“深紫外”（DUV）是一种光学方法，能够对天然和不透明表面上的单个细菌细胞进行检测和成像，包括在从厘米到微米的空间尺度上评估细菌密度和单个细胞在生物膜中的分布。DUV诱导和检测细胞或孢子固有的有机组分的天然荧光（DUV），同时避免来自底物的自发荧光干扰，使得能够在从几十厘米到微米的空间尺度上检测细菌-即，微生物和单细胞的社区。更广泛的影响这个项目从外部来源获得35万美元。预计这一技术可被广泛用作探测和测绘任何可能出现的地下偏远生命的工具，成为适用于整个深海等远端环境的“卫星成像仪”。将支持JPL/Caltech博士后在微生物生理学和荧光光谱学方面的专业知识。

Understanding the distribution of deep life in the subsurface biosphere is a major challenge to advancing our understanding of the evolution life and ecosystems on Earth. To date, all methods developed to detect microbial life in the deep biosphere involve ex-situ analysis of recovered materials from boreholes. Materials are analyzed for microbial biomass by extracting cells or cellular components, or by application of dyes to samples and performing cell counts. This latter methodology being the most standard in the industry, and represents a very time and labor intensive hands-on by eye counting of fluorescent cells under a microscope. Current methods are highly laborious and inefficient processes that involve both cell loss and the loss of information about the mineralogical context that may have influence on the microbial ecology.To meet the challenges associated with detecting and quantifying microbial life within a natural matrix, The PI requests RAPID funding to develop a novel in-situ downhole logging tool for detecting microbial life in subseafloor boreholes. "Deep UV" (DUV) is an optical method that enables detection and imaging of single bacterial cells on natural and opaque surfaces, including assessment of bacterial density and distribution of single cells to biofilms over spatial scales ranging from centimeters to microns. DUV induces and detects native fluorescence (DUV) of organic components intrinsic to the cell or spore while avoiding autofluorescence interference from the substrate, enabling detection of bacteria at spatial scales ranging from tens of centimeters to micrometers - i.e., both communities of microbes and single cells.Broader Impacts This project Leverages $350K from outside sources. It is anticipated that this technology could become widely used as a tool for detection and mapping of subsurface remote life everywhere it may occur, becoming the "satellite imager" applicable to distal environments such as represented by the entire deep sea. A JPL/Caltech post-doc with expertise in microbial physiology and fluorescence spectroscopy will be supported.