High-precision targeting workflow for volume electron microscopy

Cells are 3D objects. Therefore, volume EM (vEM) is often crucial for correct interpretation of ultrastructural data. Today, scanning EM (SEM) methods such as focused ion beam (FIB)–SEM are frequently used for vEM analyses. While they allow automated data acquisition, precise targeting of volumes of interest within a large sample remains challenging. Here, we provide a workflow to target FIB-SEM acquisition of fluorescently labeled cells or subcellular structures with micrometer precision. The strategy relies on fluorescence preservation during sample preparation and targeted trimming guided by confocal maps of the fluorescence signal in the resin block. Laser branding is used to create landmarks on the block surface to position the FIB-SEM acquisition. Using this method, we acquired volumes of specific single cells within large tissues such as 3D cultures of mouse mammary gland organoids, tracheal terminal cells in Drosophila melanogaster larvae, and ovarian follicular cells in adult Drosophila, discovering ultrastructural details that could not be appreciated before.

Quality assessment of Dobson spectrophotometers for ozone column measurements before and after automation at Arosa and Davos

The longest ozone column measurement series are based on the Dobson sun spectrophotometers developed in the 1920s by Gordon B. W. Dobson. These instruments still constitute an important part of the World Meteorological Organization's global network due to their optical qualities and ruggedness. The primary drawback of this instrument is the effort needed for its manual operation. In industrialized and some less developed countries, most stations have made the choice to replace the Dobson by the automated Brewer sun spectrophotometers, but some are still relying on the Dobson instrument. One of them is the Arosa station where both instrument types are run in parallel. Here, an automated version of the Dobson instrument was developed and implemented recently. In the present paper, the results of the analysis of simultaneous measurements from pairs of Dobson instruments that were either collocated at Arosa or Davos or operated one at each location are presented for four distinct time periods: 1992–2012 – manual vs. manual operation of collocated Dobson instruments (MMC); 2012–2013 – manual vs. automated operation of collocated Dobson instruments (MAC); 2012–2019 – automated vs. automated operation of collocated Dobson instruments (AAC); 2016–2019 – automated vs. automated operation of distant Dobson instruments (AAD). The direct comparison of two instruments using the standard operation procedure during the MMC period gives a metric necessary to validate the automated version of Dobson instruments. The direct comparison of two collocated instruments using the standard manual operation procedure reveals random differences of coincident observations with a standard deviation of ∼ 0.45 % and monthly mean differences between −1.0 % and +0.8 %. In most cases the observed biases are not statistically significant. The same analysis of two automated Dobson instruments yields significantly smaller standard deviation of ∼ 0.25 % and biases of between −0.7 % and 0.8 %. This demonstrates that the repeatability has improved with the automation, while the systematic differences are only marginally smaller. The analysis of the AAD period of coincident measurements from the distant sites Arosa and Davos reveals a small positive bias (not significant) compatible with the 250 m altitude difference. The description of the automated data acquisition and control of the Dobson instrument is presented in a separate paper (Stübi et al., 2020).

A data acquisition setup for data driven acoustic design

In this paper, we present a novel interdisciplinary approach to study the relationship between diffusive surface structures and their acoustic performance. Using computational design, surface structures are iteratively generated and 3D printed at 1:10 model scale. They originate from different fabrication typologies and are designed to have acoustic diffusion and absorption effects. An automated robotic process measures the impulse responses of these surfaces by positioning a microphone and a speaker at multiple locations. The collected data serves two purposes: first, as an exploratory catalogue of different spatio-temporal-acoustic scenarios and second, as data set for predicting the acoustic response of digitally designed surface geometries using machine learning. In this paper, we present the automated data acquisition setup, the data processing and the computational generation of diffusive surface structures. We describe first results of comparative studies of measured surface panels and conclude with steps of future research.

Quality assessment of Dobson spectrophotometers for ozone column measurements before and after automation at Arosa and Davos

The longest ozone column measurements series are based on the Dobson sun spectrophotometers developed in the 1920s by Prof. G. B. W. Dobson. These instruments still constitute an important part of the World Meteorological Organization's global network due to their optical qualities and ruggedness. The primary drawback of this instrument is the effort needed for its manual operation. In industrialized and some lesser developed countries, most stations have made the choice to replace the Dobson by the automated Brewer sun spectrophotometers but some are still relying on the Dobson instrument. One of them is the Arosa station where both instrument types are run in parallel. Here, an automated version of the Dobson instrument was developed and implemented recently. In the present paper, the results of the analysis of simultaneous measurements from pairs of Dobson instruments that were either collocated at Arosa or Davos, or operated one at each location, are presented for four distinct time periods: – 1992–2012 : Manual vs. Manual operation of Collocated Dobson instruments (MMC) – 2012–2013 : Manual vs. Automated operation of Collocated Dobson instruments (MAC) – 2012–2019 : Automated vs. Automated operation of Collocated Dobson instruments (AAC) and – 2016–2019 : Automated vs. Automated operation of Distant Dobson instruments (AAD) The direct comparison of two instruments using the standard operation procedure during the MMC period gives a metric necessary to validate the automated version of Dobson instruments. The direct comparison of two collocated instruments using the standard manual operation procedure reveals random differences of coincident observations with a standard deviation of ∼0.45 % and monthly mean differences between −1.0 and +0.8 %. In most cases the observed biases are not statistically significant. The same analysis of two automated Dobson instruments yields significantly smaller standard deviation of ∼0.25 % and biases of between −0.7 % and 0.8 %. This demonstrates that the repeatability has improved with the automation while the systematic differences are only marginally smaller. The description of the automated data acquisition and control of the Dobson instrument is presented in a separate paper (Stübi et al. , 2020).