C. elegans Synchronizer is an unique, world first, system that will allow synchronization of the nematode at 'L1'or 'Egg' stage. The CES works with a number of supporting modules, functions and protocols.

 

LabTIE C. elegans Synchronizer

 

 

The C. elegans Synchronizer (CES) is a manual worm Synchronizer that allows you to harvest small and large volumes of tight synchronized L1’s or Eggs without the use of chemicals or starvation. It enables you to get synchronized worms that do not contain phenotypes caused by bleach or food arrest. In addition the system is designed to be successfully used without extensive training and enables C. elegans experiments with healthier worms than before, giving you the freedom to focus on the experiments by eliminating the need for worm controls and variable outcomes.

     

 
Compatible with
A majority of adults and eggs in a C. elegans culture
Special features
Synchronize a L1 population without use of chemicals

The L1 C. elegans Synchronizer enables the user to manually synchronize C. elegans into a synchronous L1 population without use of chemicals.


LabTIE Services


labtie CES
 

Description

C. elegans Synchronizer

Healthy, Synhronized, Phenotype free, Results

The C. elegans Synchronizer (CES) is a manual worm Synchronizer that allows you to harvest small and large volumes of tight synchronized L1’s or Eggs without the use of chemicals or starvation. It enables you to get synchronized worms that do not contain phenotypes caused by bleach or food arrest. In addition the system is designed to be successfully used without extensive training and enables C. elegans experiments with healthier worms than before, giving you the freedom to focus on the experiments by eliminating the need for worm controls and variable outcomes.

 

 

The C. elegans Synchronizer allows you to:

  • Synchronize a mixed worm population into L1’s or Eggs
  • Obtain nematodes without phenotypes caused through bleaching
  • Obtain nematodes without phenotypes caused by starvation ( Figure 1 )
  • Start experiments without the need of additional worm controls or variable bleach outcomes
  • Higher level of synchronization (small ΔT) compared to traditional methods ( Figure 2 )
  • Harvest small as well as large volumes in a short period of time ( Figure 2 )
  • Synchronize without requiring extensive training or experience in bleaching

Benefits of using the C. elegans Synchronizer:

 

99.91% purity of L1's when using the L1 C. elegans Synchronizer system

C.elegans synchronizer puruty graphs v5

Figures: Results showing an average of 99.91% purity of L1’s when using the LabTIE L1 C. elegans Synchronizer (CES). Using the CES, the amount of L1’s present in the harvest samples compared to other stages (anomalies) was identified to determine the accuracy of the Synchronizer. A 50mL F3 generation S-medium culture was used, containing a C. elegans mixed population and was synchronized using the CES. The culture was washed for 15 minutes prior to 90 minutes of harvesting L1’s in S-media without OP50.
(A): After synchronizing, the total amount of worms were counted from 10 harvesting experiments.
(B): Anomalies were counted in each harvesting experiment. Results were plotted against the total amount of worms.
Example harvesting experiment 1: 6400 L1 worms vs 10 anomalies (7 x L2's, 3 x L3's) = 100-(100/6400*10) = 99.84% L1 purity in harvest compared to other stages present.

 

Healthier Worms
The C. elegans Synchronizer (CES) enables you to find more reliable data that is not possible with traditional synchronization methods. Variables such as temperature, time and aeration during bleaching that influence the quality and consistency of the synchronized culture are removed from the process[1]. Now you can achieve a synchronous C. elegans population without the use of chemicals that induces stress on many levels within the nematode. It is known that starvation-induced developmental arrest, a natural and drastic environmental change, leads to the generation of small RNAs that are inherited through at least three consecutive generations( Figure 1 )[2]. The effects of food arrest in relation to worm density has also been published showing change in expression in a number of genes related to metabolism and signaling[3]. Although the bleach method is still the most commonly used method for synchronization due to the lack of a better alternative, the CES is the solution that solves these issues.

 

Transgenerational Inheritance of Small RNAs Following L1 Starvation

 
Figure 1. Study showing the negative effect and phenotypes of starvation of L1’s even in F3 progeny. (A) To examine whether the small RNA response could be transmitted across generations, small RNA’s were sequenced from the fed F3 progeny of the worms that were starved as L1s. The example gene shown is (hlh−30). The P0 generation revealed that the F3 worms resemble the “starved” parents more than the “fed” parents. It was found that 52% (311/597) of the STGs that were depleted following starvation were depleted also in the F3 progeny[2]. (B) The total number of inherited STGs of different small RNA species and their overlapping targets[2].


Larger Volumes
Use any culture for synchronization, even high density liquid cultures up to 250mL can be synchronized.
Cultures containing large amounts of adults and eggs can result in a harvest of 1 million L1’s in a few hours.

 

Faster Experiments
In approximately 15 minutes, worms can be synchronized with the C. elegans Synchronizer (CES) to obtain a healthy P0 population for your experiments. Traditional methods such as bleaching has transgenerational effects on the progeny, you will have to wait multiple generations to remove phenotypes caused by transgenerational memory of previous conditions. The CES enables you to start your experiment in matter of minutes to produce healthy P0 L1 or Eggs. The conditions affecting phenotypes in progeny are often caused by chemicals and old synchronization techniques. It was found that when using Bleach or Sodium hydroxide for synchronization, transgenerational transmitted RNAs target genes with roles in nutrition are induced which were defined genes that are essential for this multigenerational effect[2]. Also, F3 offspring of starved animals due to food arrest show an increased lifespan, corroborating the notion of a transgenerational memory of past conditions.


Figure 2. Comparison of two nematode synchronizing techniques. The bleach technique was used and compared to the C. elegans Synchronizer in terms of speed and volume to be processed (yield). While bleaching takes a bit longer than the C. elegans Synchronizer to complete the protocol, the harvested L1’s showed significant difference in level of synchronization between the two techniques. Where for bleaching the difference between the fertilized eggs is about 3 a 4 hours, the
C. elegans Synchronizer was able to synchronize the L1’s within a time window of only 15 minutes.

 

 

Ordering Information

Advice:
If you want to synchronize more than 1 strain for an experiment, it is advised to order the CES with multiple Filter Sets to synchronize all required strains for your experiment. Each strain/culture can be synchronized in 15 minutes.

Example: When having 6 strains to synchronize, you would need the L1 C. elegans Synchronizer #6 (Cat.  No. CES106). The Synchronizer contains a closed loop and an absolute filter that eliminates the chance of cross contamination. Therefore the C. elegans Synchronizer does not have to be cleaned (CIP) in between synchronizing runs.

Product

Stabilization Filter

Harvest Filter*

Cat. No.

L1 C. elegans Synchronizer #1

1

1

CES101

L1 C. elegans Synchronizer #6

2

6

CES106

Egg C. elegans Synchronizer #1

1

1

CES201

Egg C. elegans Synchronizer #6

2

6

CES206

L1 and Egg C. elegans Synchronizer Combo System #1

1

1

CES301

L1 and Egg C. elegans Synchronizer Combo System #6

2

6

CES306

Freeze-dried OP50 – 40 vials

-

-

1.17.040

*To allow synchronization of different C. elegans strains an additional Harvest filter is needed. Both filters can be re-used indefinitely.

 

Learn more at labtie.com. Or contact us at

 

References

  1. Porta-de-la-Riva, M., Fontrodona, L., Villanueva, A., Cerón, J. Basic Caenorhabditis elegans Methods: Synchronization and Observation. J. Vis. Exp. (64), e4019 10.3791/4019, DOI : 10.3791/4019 (2012)
  2. Rechavi, O., Houri-Ze’evi, L., Anava, S., Sho Goh, W. S., Kerk, S. Y., Hannon, G. J., & Hobert, O. (2014). Starvation-Induced Transgenerational Inheritance of Small RNAs in C. elegans. Cell, 158(2), 277–287. http://doi.org/10.1016/j.cell.2014.06.020
  3. Rando, Oliver J., Io Long Chan, and Colin Conine. "Effects of larval density on gene regulation in C. elegans during routine L1 synchronization." bioRxiv(2018): 284927.

 

In Development
 
Compatible with
A majority of adults and eggs in a C. elegans culture
Special features
Synchronize an egg population without use of chemicals

The Egg C. elegans Synchronizer enables the user to manually synchronize C. elegans into a synchronous egg population without use of chemicals.


LabTIE Services


labtie ces diagram
 

Description

Get Sychronizer Egg's without Bleach or Food arrest in Five steps

C. elegans Synchronizer

The C. elegans Synchronizer (CES) is a manual worm Synchronizer that allows you to harvest small and large volumes of tight synchronized L1’s or Eggs without the use of chemicals or starvation. It enables you to get synchronized worms that do not contain phenotypes caused by bleach or food arrest. In addition the system is designed to be successfully used without extensive training and enables C. elegans experiments with healthier worms than before, giving you the freedom to focus on the experiments by eliminating the need for worm controls and variable outcomes.

 

 

The C. elegans Synchronizer allows you to:

  • Synchronize a mixed worm population into L1’s or Eggs
  • Obtain nematodes without phenotypes caused through bleaching
  • Obtain nematodes without phenotypes caused by starvation ( Figure 1 )
  • Start experiments without the need of additional worm controls or variable bleach outcomes
  • Higher level of synchronization (small ΔT) compared to traditional methods ( Figure 2 )
  • Harvest small as well as large volumes in a short period of time ( Figure 2 )
  • Synchronize without requiring extensive training or experience in bleaching

Benefits of using the C. elegans Synchronizer:

 

Healthier Worms
The C. elegans Synchronizer (CES) enables you to find more reliable data that is not possible with traditional synchronization methods. Variables such as temperature, time and aeration during bleaching that influence the quality and consistency of the synchronized culture are removed from the process[1]. Now you can achieve a synchronous C. elegans population without the use of chemicals that induces stress on many levels within the nematode. It is known that starvation-induced developmental arrest, a natural and drastic environmental change, leads to the generation of small RNAs that are inherited through at least three consecutive generations( Figure 1 )[2]. The effects of food arrest in relation to worm density has also been published showing change in expression in a number of genes related to metabolism and signaling[3]. Although the bleach method is still the most commonly used method for synchronization due to the lack of a better alternative, the CES is the solution that solves these issues.

 

Transgenerational Inheritance of Small RNAs Following L1 Starvation

 
Figure 1. Study showing the negative effect and phenotypes of starvation of L1’s even in F3 progeny. (A) To examine whether the small RNA response could be transmitted across generations, small RNA’s were sequenced from the fed F3 progeny of the worms that were starved as L1s. The example gene shown is (hlh−30). The P0 generation revealed that the F3 worms resemble the “starved” parents more than the “fed” parents. It was found that 52% (311/597) of the STGs that were depleted following starvation were depleted also in the F3 progeny[2]. (B) The total number of inherited STGs of different small RNA species and their overlapping targets[2].


Larger Volumes
Use any culture for synchronization, even high density liquid cultures up to 250mL can be synchronized.
Cultures containing large amounts of adults and eggs can result in a harvest of 100k egg’s in a few hours.

 

Faster Experiments
In approximately 30 minutes, eggs can be synchronized with the C. elegans Synchronizer (CES) to obtain a healthy egg population for your experiments. Traditional methods such as bleaching has transgenerational effects on the progeny, you will have to wait multiple generations to remove phenotypes caused by transgenerational memory of previous conditions. The CES enables you to start your experiment in matter of minutes to produce healthy P0 L1 or Eggs. The conditions affecting phenotypes in progeny are often caused by chemicals and old synchronization techniques. It was found that when using Bleach or Sodium hydroxide for synchronization, transgenerational transmitted RNAs target genes with roles in nutrition are induced which were defined genes that are essential for this multigenerational effect[2]. Also, F3 offspring of starved animals due to food arrest show an increased lifespan, corroborating the notion of a transgenerational memory of past conditions.


Figure 2. Comparison of two nematode synchronizing techniques. The bleach technique was used and compared to the C. elegans Synchronizer in terms of speed and volume to be processed (yield). While bleaching takes a bit longer than the C. elegans Synchronizer to complete the protocol, the harvested L1’s showed significant difference in level of synchronization between the two techniques. Where for bleaching the difference between the fertilized eggs is about 3 a 4 hours, the
C. elegans Synchronizer was able to synchronize the L1’s within a time window of only 15 minutes.

 

 

Ordering Information

Advice:
If you want to synchronize more than 1 strain for an experiment, it is advised to order the CES with multiple Filter Sets to synchronize all required strains for your experiment. Each strain/culture can be synchronized in 15 minutes.

Example: When having 6 strains to synchronize, you would need the L1 C. elegans Synchronizer #6 (Cat.  No. CES106). The Synchronizer contains a closed loop and an absolute filter that eliminates the chance of cross contamination. Therefore the C. elegans Synchronizer does not have to be cleaned (CIP) in between synchronizing runs.

Product

Stabilization Filter

Harvest Filter*

Cat. No.

L1 C. elegans Synchronizer #1

1

1

CES101

L1 C. elegans Synchronizer #6

2

6

CES106

L1 C. elegans Synchronizer #12

2

12

CES112

Egg C. elegans Synchronizer #1

1

1

CES201

Egg C. elegans Synchronizer #6

2

6

CES206

Egg C. elegans Synchronizer #12

2

12

CES212

L1 and Egg C. elegans Synchronizer Combo System #1

1

1

CES301

L1 and Egg C. elegans Synchronizer Combo System #6

2

6

CES306

L1 and Egg C. elegans Synchronizer Combo System #12

2

12

CES312

Synchronizer Silicone Tubing set (re-usable)

-

-

CES190

Freeze-dried OP50 – 40 vials

-

-

1.17.040

*To allow synchronization of different C. elegans strains an additional Harvest filter is needed. Both filters can be re-used indefinitely.

 

Learn more at labtie.com. Or contact us at

 

References

  1. Porta-de-la-Riva, M., Fontrodona, L., Villanueva, A., Cerón, J. Basic Caenorhabditis elegans Methods: Synchronization and Observation. J. Vis. Exp. (64), e4019 10.3791/4019, DOI : 10.3791/4019 (2012)
  2. Rechavi, O., Houri-Ze’evi, L., Anava, S., Sho Goh, W. S., Kerk, S. Y., Hannon, G. J., & Hobert, O. (2014). Starvation-Induced Transgenerational Inheritance of Small RNAs in C. elegans. Cell, 158(2), 277–287. http://doi.org/10.1016/j.cell.2014.06.020
  3. Rando, Oliver J., Io Long Chan, and Colin Conine. "Effects of larval density on gene regulation in C. elegans during routine L1 synchronization." bioRxiv(2018): 284927.