A single fixation protocol for proteome-wide immunofluorescence localization studies

2010 ◽  
Vol 73 (6) ◽  
pp. 1067-1078 ◽  
Author(s):  
Charlotte Stadler ◽  
Marie Skogs ◽  
Hjalmar Brismar ◽  
Mathias Uhlén ◽  
Emma Lundberg
Keyword(s):  
Immunofluorescence Localization ◽  
Fixation Protocol ◽  
Single Fixation

Detecting single-fixation changes in pictures

1997 ◽  
Author(s):  
Christopher B. Currie ◽  
George W. McConkle
Keyword(s):  
Single Fixation

scholarly journals Depletion of Centromeric MCAK Leads to Chromosome Congression and Segregation Defects Due to Improper Kinetochore Attachments

2004 ◽  
Vol 15 (3) ◽  
pp. 1146-1159 ◽  
Author(s):  
Susan L. Kline-Smith ◽  
Alexey Khodjakov ◽  
Polla Hergert ◽  
Claire E. Walczak
Keyword(s):  
Essential Role ◽  
Dominant Negative ◽  
Primary Role ◽  
Complex Behavior ◽  
Chromosome Movement ◽  
Immunofluorescence Localization ◽  
Microtubule Attachment ◽  
Chromosome Congression

The complex behavior of chromosomes during mitosis is accomplished by precise binding and highly regulated polymerization dynamics of kinetochore microtubules. Previous studies have implicated Kin Is, unique kinesins that depolymerize microtubules, in regulating chromosome positioning. We have characterized the immunofluorescence localization of centromere-bound MCAK and found that MCAK localized to inner kinetochores during prophase but was predominantly centromeric by metaphase. Interestingly, MCAK accumulated at leading kinetochores during congression but not during segregation. We tested the consequences of MCAK disruption by injecting a centromere dominant-negative protein into prophase cells. Depletion of centromeric MCAK led to reduced centromere stretch, delayed chromosome congression, alignment defects, and severe missegregation of chromosomes. Rates of chromosome movement were unchanged, suggesting that the primary role of MCAK is not to move chromosomes. Furthermore, we found that disruption of MCAK leads to multiple kinetochore–microtubule attachment defects, including merotelic, syntelic, and combined merotelic-syntelic attachments. These findings reveal an essential role for Kin Is in prevention and/or correction of improper kinetochore–microtubule attachments.

scholarly journals Optimal observer model of single-fixation oddity search predicts a shallow set-size function

2007 ◽  
Vol 7 (10) ◽  
pp. 1 ◽  
Author(s):  
Wade Schoonveld ◽  
Steve S. Shimozaki ◽  
Miguel P. Eckstein
Keyword(s):  
Set Size ◽  
Size Function ◽  
Observer Model ◽  
Single Fixation

Efficacy and safety of a collagen matrix for cranial and spinal dural reconstruction using different fixation techniques

2008 ◽  
Vol 109 (2) ◽  
pp. 215-221 ◽  
Author(s):  
Ruediger Stendel ◽  
Marco Danne ◽  
Ingo Fiss ◽  
Ilse Klein ◽  
Andreas Schilling ◽  
...  
Keyword(s):  
Collagen Matrix ◽  
Deep Infection ◽  
Surgical Revision ◽  
Neurosurgical Procedures ◽  
Efficacy And Safety ◽  
Local Skin ◽  
Dural Graft ◽  
Csf Leakage ◽  
Fixation Techniques ◽  
Single Fixation

Object The use of dural grafts is frequently unavoidable when tension-free dural closure cannot be achieved following neurosurgical procedures or trauma. Biodegradable collagen matrices serve as a scaffold for the regrowth of natural tissue and require no suturing. The aim of this study was to investigate the efficacy and safety of dural repair with a collagen matrix using different fixation techniques. Methods A total of 221 patients (98 male and 123 female; mean age 55.6 ± 17.8 years) undergoing cranial (86.4%) or spinal (13.6%) procedures with the use of a collagen matrix dural graft were included in this retrospective study. The indications for use, fixation techniques, and associated complications were recorded. Results There were no complications of the dural graft in spinal use. Five (2.6%) of 191 patients undergoing cranial procedures developed infections, 3 of which (1.6%) were deep infections requiring surgical revision. There was no statistically significant relationship between the operative field status before surgery and the occurrence of a postoperative wound infection (p = 0.684). In the 191 patients undergoing a cranial procedure, cerebrospinal fluid (CSF) collection occurred in 5 patients (2.6%) and a CSF fistula in 5 (2.6%), 3 of whom (1.6%) required surgical revision. No patient who underwent an operation with preexisting CSF leakage had postoperative CSF leakage. Postoperative infection significantly increased the risk for postoperative CSF leakage. The collagen matrix was used without additional fixation in 124 patients (56.1%), with single fixation in 55 (24.9%), and with multiple fixations in 42 (19%). There were no systemic allergic reactions or local skin changes. Follow-up imaging in 112 patients (50.7%) revealed no evidence of any adverse reaction to the collagen graft. Conclusions The collagen matrix is an effective and safe cranial and spinal dural substitute that can be used even in cases of an existing local infection. Postoperative deep infection increases the risk for CSF leakage.

scholarly journals A perfusion-fixation procedure for the concurrent demonstration of Timm's, horseradish peroxidase (HRP), and acethycholinesterase (AChE) histochemistry.

1984 ◽  
Vol 32 (10) ◽  
pp. 1113-1116 ◽  
Author(s):  
M B Moss ◽  
D L Rosene
Keyword(s):  
Horseradish Peroxidase ◽  
Sodium Sulfide ◽  
Prolonged Exposure ◽  
Initial Exposure ◽  
Sodium Sulfide Solution ◽  
Acetylcholinesterase Histochemistry ◽  
The Central Nervous System ◽  
Fixation Protocol ◽  
Perfusion Fixation ◽  
Sulfide Solution

The sulfide-silver method of Timm has been a widely used histochemical technique to demonstrate the presence of heavy metals in biological tissue, particularly in the central nervous system. However, the use of this method or its several modifications results in less than optimal morphological preservation and requires embedding the tissue in paraffin or freezing it and cutting it directly onto slides with a cryostat. These procedures can decrease the sensitivity and limit the application of other histochemical procedures, particularly when experiments necessitate processing large specimens or reaction procedures require techniques using free-floating sections. A perfusion-fixation protocol is described that yields sufficient fixation to cut whole frozen blocks of tissue with a sliding microtome, permits the use of free-floating sections, and allows the concurrent demonstration of horseradish peroxidase and acetylcholinesterase histochemistry without loss of sensitivity. The method consists of a short initial exposure to a sodium sulfide solution followed by a prolonged exposure to a combined sulfide-aldehyde fixative solution.

Eye movement control in Turkish sentence reading

2020 ◽  
pp. 174702182096331
Author(s):  
Ayşegül Özkan ◽  
Figen Beken Fikri ◽  
Bilal Kırkıcı ◽  
Reinhold Kliegl ◽  
Cengiz Acartürk
Keyword(s):  
Wide Spectrum ◽  
Distributed Processing ◽  
Movement Control ◽  
Predictive Processing ◽  
Base Line ◽  
Discourse Comprehension ◽  
Morphological Effect ◽  
Fixation Durations ◽  
Single Fixation ◽  
High Level

Reading requires the assembly of cognitive processes across a wide spectrum from low-level visual perception to high-level discourse comprehension. One approach of unravelling the dynamics associated with these processes is to determine how eye movements are influenced by the characteristics of the text, in particular which features of the words within the perceptual span maximise the information intake due to foveal, spillover, parafoveal, and predictive processing. One way to test the generalisability of current proposals of such distributed processing is to examine them across different languages. For Turkish, an agglutinative language with a shallow orthography–phonology mapping, we replicate the well-known canonical main effects of frequency and predictability of the fixated word as well as effects of incoming saccade amplitude and fixation location within the word on single-fixation durations with data from 35 adults reading 120 nine-word sentences. Evidence for previously reported effects of the characteristics of neighbouring words and interactions was mixed. There was no evidence for the expected Turkish-specific morphological effect of the number of inflectional suffixes on single-fixation durations. To control for word-selection bias associated with single-fixation durations, we also tested effects on word skipping, single-fixation, and multiple-fixation cases with a base-line category logit model, assuming an increase of difficulty for an increase in the number of fixations. With this model, significant effects of word characteristics and number of inflectional suffixes of foveal word on probabilities of the number of fixations were observed, while the effects of the characteristics of neighbouring words and interactions were mixed.

Visual Cortex Neurons of Monkeys and Cats: Temporal Dynamics of the Spatial Frequency Response Function

2004 ◽  
Vol 91 (6) ◽  
pp. 2607-2627 ◽  
Author(s):  
Robert A. Frazor ◽  
Duane G. Albrecht ◽  
Wilson S. Geisler ◽  
Alison M. Crane
Keyword(s):  
Spatial Frequency ◽  
Striate Cortex ◽  
Temporal Dynamics ◽  
Temporal Integration ◽  
Direction Selectivity ◽  
Response Latencies ◽  
Low Frequencies ◽  
Transient Nature ◽  
Integration Strategies ◽  
Single Fixation

We measured the responses of striate cortex neurons as a function of spatial frequency on a fine time scale, over the course of an interval that is comparable to the duration of a single fixation (200 ms). Stationary gratings were flashed on for 200 ms and then off for 300 ms; the responses were analyzed at sequential 1-ms intervals. We found that 1) the preferred spatial frequency shifts through time from low frequencies to high frequencies, 2) the latency of the response increases as a function of spatial frequency, and 3) the poststimulus time histograms (PSTHs) are relatively shape-invariant across spatial frequency. The dynamic shifts in preferred spatial frequency appear to be a simple consequence of the latency shifts and the transient nature of the PSTH. The effects of these dynamic shifts on the coding of spatial frequency information are examined within the context of several different temporal integration strategies, and pattern-detection performance is determined as a function of the interval of integration, following response onset. The findings are considered within the context of related investigations as well as a number of functional issues: motion selectivity in depth, “coarse-to-fine” processing, direction selectivity, latency as a code for stimulus attributes, and behavioral response latency. Finally, we demonstrate that the results are qualitatively consistent with a simple feedforward model, similar to the one originally proposed in 1962 by Hubel and Wiesel, that incorporates measured differences in the response latencies and the receptive field sizes of different lateral geniculate nucleus inputs.

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