Difference between revisions of "O2k-Titration Set"

Latest revision as of 17:46, 11 October 2022

O2k-Titration Set

MitoPedia O2k and high-resolution respirometry: O2k-Open Support

Description

The O2k-Titration Set consists of Hamilton microsyringes (6 x 10 mm³ and 3 spare plungers, 6 x 25 mm³, 1 x 50 mm³, 1 x 100 mm³, 1 x 500 mm³; fixed needles with rounded tips), provided in the Syringe Storage Box with Syringe Labels, a set of two Syringe Racks with Syringe Collars, and a set of two Tube Racks.

Reference: Product details and purchasing information

Additional resources

»MiPNet09.12 O2k-Titrations

»MiPNet19.14 SOP Hamilton microsyringes

Note: Titration of chemicals in respirometry

Overshoots or short interruptions of the flux after the injection of chemicals do not have a meaning in respirometric measurements. A disturbance of the signal is only observed due to the injected chemical and does not represent mitochondrial respiration. With the addition of a chemical solved in ethanol a little bit of oxygen is always introduced, since ethanol has higher oxygen solubility than water.

After a titration, it is common to observe liquid accumulated on top of the stopper receptacle. When this happens, we recommend removing this excess of liquid as it can lead to O₂ backdiffusion into the chamber. For this, we recommend using the ISS, placing the tip close to the capillary, but not directly on top of it.

Further information

Safety information: fixed needles are used with rounded tips, not usable for injection into tissues.
Hamilton microsyringes are specifically produced for O2k-titrations and can be ordered directly from Oroboros Instruments to assure exact fitting with the O2k-Stopper and O2k-Chamber dimensions.
Visit also: Hamilton.
Eliminate rough edges with 3M™ Wetordry™ Paper Sheet P1000 by gentle rubbing.
Use a cleaning wire for the needle. If a syringe is clogged , sometimes it is also helpful to fill it from the back with EtOH or H₂O and try to press the plunger gently down.

MitoPedia O2k and high-resolution respirometry: O2k-Open Support, O2k-Respirometry, O2k-FluoRespirometry

@@ Line 1: / Line 1: @@
-{{Product
+{{Technical support}}
-|description=The '''Titration Set''' consists of 6x 10 µl, 6x 25 µl, and 2x 50 µl Hamilton glass syringes, and 3 spare plungers for the 10 µl syringes. It is a basic HRR-Accessory included in the [[O2k-Core]] Unit.
+__TOC__
-|product id=20820-01
+{{MitoPedia without banner
-|product type=O2k, O2k-Module, O2k-Core, Microsyringe, Catalogue
+|abbr=
-|info=[[MiPNet09.12 O2k-Titrations]]
+|description=[[Image:O2k-Titration Set.JPG|right|180px]] The '''O2k-Titration Set''' consists of Hamilton microsyringes (6 x 10 mm<sup>3</sup> and 3 spare plungers, 6 x 25 mm<sup>3</sup>, 1 x 50 mm<sup>3</sup>, 1 x 100 mm<sup>3</sup>, 1 x 500 mm<sup>3</sup>; fixed needles with rounded tips), provided in the [[Syringe Storage Box]] with [[Syringe Labels]], a set of two [[Syringe Racks]] with [[Syringe Collars]], and a set of two [[Tube Racks]].
-|product image=[[Image:Hamilton Syringes for Manual Titration.jpg|right|180px]]
+|info= [https://www.oroboros.at/index.php/product/o2k-titration-set/ '''Product details and purchasing information''']
 }}
-== The '''Titration Set''' ''consists of'' ==
-:6 [[Microsyringe\10 mm3 51/0.13 mm]] 10 mm<sup>3</sup>
+== Additional resources ==
-:6 [[Microsyringe\25 mm3 51/0.15 mm]] 25 mm<sup>3</sup>
+:::: »[[MiPNet09.12 O2k-Titrations]] </br>
-:2 [[Microsyringe\50 mm3 51/0.15 mm]] 50 mm<sup>3</sup>
+:::: »[[MiPNet19.14 SOP Hamilton microsyringes]]
-:3 [[Plunger\10 mm3]] for 10 mm<sup>3</sup> syringe
-== '''Handling of the syringes''' ==
+== Note: Titration of chemicals in respirometry ==
-===General information===
+:::: Overshoots or short interruptions of the flux after the injection of chemicals do not have a meaning in respirometric measurements. A disturbance of the signal is only observed due to the injected chemical and does not represent mitochondrial respiration. With the addition of a chemical solved in ethanol a little bit of oxygen is always introduced, since ethanol has higher oxygen solubility than water.
-Use separate syringes for substrates, uncoupler and inhibitors.
-Colour code of syringes:
+::::  After a titration, it is common to observe liquid accumulated on top of the stopper receptacle. When this happens, we recommend removing this excess of liquid as it can lead to O<sub>2</sub> backdiffusion into the chamber. For this, we recommend using the [[ISS]], placing the tip close to the capillary, but not directly on top of it.
-* Substrate: white
-* Uncoupler: blue
-* Inhibitors: red
-* Digitonin: yellow
-===Preparation of syringes before experiment===
+=== Further information ===
-* Prepare syringes according to [[SUIT protocol]] and [[MiPNet09.12 O2k-Titrations | O2k-Titrations]].
+:::* Safety information: fixed needles are used with rounded tips, not usable for injection into tissues.
-* Prepare a rack with three 50 ml falcons for washing procedure:
+:::* Hamilton microsyringes are specifically produced for O2k-titrations and can be ordered directly from Oroboros Instruments to assure exact fitting with the O2k-Stopper and O2k-Chamber dimensions.
-:* First rinse (distilled water)
+:::* Visit also: [http://www.hamiltoncompany.com/products/syringes-and-needles Hamilton].
-:* Distilled water
+:::* Eliminate rough edges with 3M™ Wetordry™ Paper Sheet P1000 by gentle rubbing.
-:* 100% EtOH
+:::* Use a cleaning wire for the needle. If a syringe is clogged , sometimes it is also helpful to fill it from the back with EtOH or H<sub>2</sub>O and try to press the plunger gently down.
-:* Prepare beaker for waste
-* Washing procedure:
-:* Discard residual chemical into the beaker
-:* Plunge syringe in 'first rinse' falcon (distilled water) and wipe it with a small kimwipe. Take care to rinse not only the needle itself but also the needle base
-:* Wash syringe three times in falcon filled with distilled water or 100% EtOH (depends on whether reagents were dissolved in distilled water or EtOH).
-:* Syringes used for AmR or CCCP: wash twice with DMSO (flush syringe surface with distilled water between washing steps).
-===Syringes in use===
-* Hold the syringe on the top to prevent warming of the glass.
-* Fill the syringe above desired mark in order to have a little bit of excess.
-* Take care not to produce bubbles when filling the syringe – this causes inaccuracies in measurement.
-* Fill the syringe not too fast but steady. Pumping is also ok.
-* Before titrating the chemical into the chamber: carefully press plunger and check if a small drop appears on top of the needle
-* Be swift when titrating in the chamber, especially when titrating EtOH due to its volatile nature.
-* After titrating in the chamber, discard the residual chemical into the beaker and rinse the outside of the syringe with distilled water (plunging in the falcon labelled 'first rinse') and dab it gently using small kimwipes.
-* Store syringes in 15 ml glass epprouvettes during the experiment in used solvent (distilled water or EtOH, use a separate epprouvette for every syringe).
-* Caution: take care to specifically rinse the syringe used for ADP (with MgCl2) IMMEDIATELY after use (on the surface but more importantly inside 3 times with distilled water) as a precipitate may form and cause blocking of the needle.
-===Preparation of syringes after experiment and storage===
+{{MitoPedia O2k and high-resolution respirometry
-* Take care of the washing order: first substrate-, then uncoupler- and finally inhibitor syringes.
+|mitopedia O2k and high-resolution respirometry=O2k-Open Support, O2k-Respirometry, O2k-FluoRespirometry
-* Wash three times with last used solvent, take care again to rinse also the needle base, rinse three times with EtOH 100% (use a small beaker).
+}}
-* Store syringes in dry condition.
-<u>Note:</u><br>
-When using a syringe with a chemical different from the usually assigned one then a washing step with distilled water is crucial to prevent medium contamination by carry-over!
-=== Further information ===
-* [[MiPNet19.14 SOP Hamilton microsyringes | MiPNet19.14]] - SOP for manual O2k-titrations with Hamilton microsyringes
-* Visit also: [http://www.hamiltoncompany.com/Syringes/ Hamilton] and [http://www.hamiltoncompany.com/Syringes/techInfo/PDFs/L20058_care_use_guide.pdf Hamilton info pdf]
-* Eliminate rugh edges with 3M™ Wetordry™ Paper Sheet P1000 by gentle rubbing
-* Use a cleaning wire for the needle. If a syringe is clocked , sometimes it is also helpful to fill it from the back with EtOH or H2O and try to press the plunger gentle down.