Coating particles with dextran-conjugated fluorescent dyes or other hydrophilic compounds.

نویسندگان

  • Josh L Morgan
  • Daniel Kerschensteiner
چکیده

Imaging and reconstruction of developing neurons require cells that are labeled in a way that distinguishes them from their neighbors. This can be achieved with ballistic labeling, which refers to the delivery of a cell label by means of carrier particles (tungsten or gold) propelled from a pressurized gun. Ballistic delivery can reach many dispersed cells in one shot and can deploy a wide variety of cell markers to neurons in diverse preparations. The three most commonly used types of ballistic labels are carbocyanine dyes, dextran-conjugated fluorescent markers, and DNA plasmids. The primary advantage of ballistic labeling is that multiple dispersed cells can be labeled quickly in live or fixed tissue. This article describes a protocol for coating tungsten particles with dextran-conjugated fluorescent dyes or ion indicators. Such hydrophilic compounds conjugated to dextran are water soluble, and therefore they are excellent indicators for functional studies within living cells. This protocol was developed for labeling ganglion cells in retinal flat mounts.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Fourier transformed spectral bio-imaging for studying the intracellular fate of liposomes.

BACKGROUND To improve the efficiency of liposomal drug targeting systems, it is necessary to understand the mechanism of liposome uptake by the cell and to follow the intracellular fate of internalized liposomes and their contents. METHODS We applied multiple-color fluorescence imaging spectroscopy, using a combination of five fluorescent dyes with a significant spectral overlap. pH-sensitive...

متن کامل

Effective permeability of hydrophilic substances through walls of lymph vessels: roles of endothelial barrier.

The wall effective permeability of hydrophilic substances labeled with fluorescent dyes was evaluated in an isolated cannulated rat single lymph vessel through a videomicroscope system. Sodium fluorescein (NaFl; 332 mol wt) and FITC-dextrans (4,400, 12,000, and 71,200 mol wt) were administered into the intraluminal space of the lymph vessels and then excited by a Xenon lamp. Changes in the fluo...

متن کامل

Phenylenevinylene conjugated oligoelectrolytes as fluorescent dyes for mammalian cell imaging.

Conjugated phenylenevinylene oligoelectrolytes, which consist of a phenylenevinylene core equipped at each end with hydrophilic pendent groups, are shown to be good candidates for mammalian cell membrane staining. When used in the micromolar concentration range, they express low to moderate cell toxicity for selected regular and cancerous cell lines as tested for adherent and suspension cells.

متن کامل

Synthesis and photophysical properties of pyrene-based green fluorescent dyes: butterfly-shaped architectures.

A few pyrene-based fluorescent compounds were synthesized using Pd/Cu-catalyzed cross-coupling reaction. Photophysical properties of the π-conjugated pyrene derivatives were studied and the results indicate materials with high quantum efficiency and high extinction coefficient. No π stacking was observed in the crystal lattice. The molecules described here may be useful in developing sensors or...

متن کامل

14.6 FluoSpheres and TransFluoSpheres Microspheres for Tracing

FluoSpheres and TransFluoSpheres polystyrene microspheres 1 satisfy several prerequisites of ideal long-term biological tracers. Because the dyes in our microspheres are incorporated throughout the microsphere rather than just on its surface, the fluorescence output per microsphere is significantly greater than that obtained from protein or dextran conjugates (Table 6.6) and is relatively immun...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Cold Spring Harbor protocols

دوره 2012 1  شماره 

صفحات  -

تاریخ انتشار 2012