Hello, I am Jan Dolzer, product manager for the Sutter Instrument P-2000 Laser-based Micropipette Puller.
We are producing a series of instructional videos that demonstrate
how individual parameters of the pull program impact the shape of the final pipette.
Our ultimate goal is to give you an in-depth understanding of how each of the parameters affects the geometry
of the pipette, and what you need to change if you want to achieve, say,
a shorter taper or a smaller electrode opening.
This introductory video will give you a little general background about the P-2000 Puller
and will help us understand what we see in those close-up videos.
Let us take a look at the instrument.
Under the tinted lid, the two puller bars hold and position the glass capillary.
The shroud between the puller bars encloses the retro mirror, which redirects
and focuses the laser beam to achieve even heating of the glass capillary.
In our first set of video clips, we used 1-mm thick-wall quartz glass, item number Q100-50-10.
Sample program number 1 pulls a patch pipette with a short, 4-mm taper and a pipette opening
of about 1 micrometer in diameter.
We are loading the quartz glass capillary into the puller bar on one side,
bring the two puller bars together, and clamp the other side.
The lid must be closed, Program # 1 is selected, and: ACTION!
Since you are watching this video, you have probably done this two or three times before.
Therefore, let's now move into our laser safety lab
and take a look at things the way you have not seen them yet.
In our series of tutorial videos,
we will watch micropipettes in the process of being pulled from premier seating.
Viewing the P-2000 Puller from its back at an angle, the retro mirror is oriented like this.
Zooming in from this perspective gives us an example of the Diagonal View, in which we get a good look
of the retro mirror, the clamps of the puller bars and, of course, the glass capillary.
We will take a closer look from the same point of view, focusing on the glass capillary.
Here this sequence again in slow motion.
Quartz glass lights up when it is hot enough to be pulled, thus allowing us to easily monitor the heat distribution.
Note how the hourglass is formed during the first heating cycle,
and the glass separates very quickly during the second cycle.
In another point of view we use in our videos, the camera is positioned directly behind the retro mirror,
just above the axis of the laser beam. This is referred to as the Parallel View.
Most of the time in parallel view, we will have the lights dimmed.
This gives us a higher-quality image of the heat distribution within the quartz capillary.
The main objective of our tutorial videos is, of course, teaching you
how to create your own pipette programs.
In one of the first scenarios, we will make a copy of Program #1,
which pulls patch pipettes, to Program #71, and then vary parameters, such as the velocity.
Then we will discuss the impact of different velocities on the shape of the final pipette.
In the example shown here, the length of the hourglass increases with the velocity.
An important factor in being proficient at pulling pipettes is the ability to recognize
and correctly interpret the symptoms of when a parameter has been pushed beyond its useful range
for a certain type of glass.
To learn this, we will take close-up looks at pipette tips pulled with appropriate parameter sets
and compare them to pipettes pulled with parameters outside the reasonable range.
Let's finish with a word of caution.
The P-2000 Puller is a Class 1 laser product, which means in everyday operation,
the laser radiation is safely contained inside the enclosed housing.
Redundant interlocks and other safety features reliably protect the user from exposure to the laser beam.
To show pipettes while they are being pulled, we have produced the detail images in our videos
using a specially modified instrument running under controlled conditions in our laser safety lab.
The 10-W CO2 laser in the P-2000 Puller produces an invisible beam that is powerful enough
to melt quartz glass. That means it is easily strong enough
to cause serious burns to your skin and, especially, to your cornea.
We don't want you to get hurt. So please do not disable the safety features on your puller at home!
What you should try at home, however, is watching our tutorial videos
and experimenting with the program parameters yourself.
A good microscope with long-distance optics helps tremendously monitoring the effect of a parameter change.
If you need tech support, go to www.sutter.com or give us a call at +1 415-883-0128.
Enjoy our videos!