Subsurface Protein Controls
Subsurface Protein Controls - Related Videos
11:34
Subsurface Defect Localization by Structured Heating Using Laser Projected Photothermal Thermography
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Cited by 7 •
2017
This method aims at locating vertical subsurface defects. Here, we couple a laser with a spatial light modulator and trigger its video input to heat a sample surface deterministically with two anti-phased modulated lines while acquiring highly resolved thermal images. The defect position is retrieved from evaluating thermal wave interference minima.
07:58
Data Processing Methods for 3D Seismic Imaging of Subsurface Volcanoes: Applications to the Tarim Flood Basalt
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Cited by 3 •
2017
Three-dimensional (3D) reflection seismology is a powerful method for imaging subsurface volcanoes. By using industrial 3D seismological data from the Tarim Basin, we illustrate how to extract the sills and the conduits of subsurface volcanoes from seismic data cubes.
13:27
Exploring the Effects of Atmospheric Forcings on Evaporation: Experimental Integration of the Atmospheric Boundary Layer and Shallow Subsurface
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Cited by 9 •
2015
A protocol for the design and construction of a soil tank interfaced to a small climate controlled wind tunnel to study the effects of atmospheric forcings on evaporation is presented. Both the soil tank and wind tunnel are instrumented with sensor technologies for the continuous in situ measurement of environmental conditions.
06:09
Studying the Protein Quality Control System of D. discoideum Using Temperature-controlled Live Cell Imaging
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2016
The social amoebae Dictyostelium discoideum has recently been established as a system to study protein misfolding and proteostasis. Here, we describe a new imaging-based methodology to study temperature-induced protein aggregation and the cellular stress response in D. discoideum.
10:47
Immuno-fluorescence Assay of Leptospiral Surface-exposed Proteins
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Cited by 23 •
2011
An efficient method to assess surface-exposure of leptospiral proteins is described. The method is specifically designed to avoid disruption of the fragile outer membrane of leptospiral cells. This technique requires employment of several negative controls to assess the integrity of the outer membrane and specificity of antibody reaction.
FAQs
The key comparison is probe binding before and after access to the cell interior is altered. In intact cells, the microbial envelope can restrict antibodies or probes from reaching subsurface targets. After permeabilization or disruption, those targets become accessible. A marked increase in binding after this treatment supports internal localization, whereas similar binding in both conditions suggests exterior accessibility.
Surface markers provide a reference for whether the experimental comparison preserves the expected distinction between accessible and restricted targets. Their binding in intact cells helps identify exterior components, while the subsurface signal is evaluated separately after permeabilization or disruption. This comparison strengthens localization conclusions by showing that probe access, rather than an unexplained loss of recognition, accounts for the result.
Negative controls establish the background level of probe binding that is unrelated to the microbial protein of interest. They are especially important when comparing intact and treated cells, because handling or disruption could affect nonspecific interactions. Interpreting the experimental signal against this background helps investigators decide whether increased binding after treatment reflects newly accessible subsurface antigen rather than nonspecific probe association.
Investigators first examine probe or antibody binding in intact microbial cells, then repeat the assessment after permeabilization or disruption. They include surface markers to monitor exterior accessibility and negative controls to identify background binding. Comparing the resulting signals allows the experimenter to determine whether recognition is already possible at the surface or emerges only when internal targets become reachable.
They are useful when protein localization affects how a host immune system can recognize a microbe. An antigen beneath the cell surface may be present but inaccessible to antibodies or other probes under intact-cell conditions. Establishing that distinction helps researchers interpret immune-recognition experiments and avoid treating hidden proteins as though they were directly exposed during host-pathogen interactions.
Localization results can inform the evaluation of diagnostic targets, vaccine candidates, and microbial mechanisms that depend on host access to proteins. A protein that becomes detectable only after permeabilization may have different relevance from one recognized on intact cells. These controls therefore connect experimental binding patterns with practical questions about antigen accessibility, immune recognition, and infection biology.